Expression and polymorphism of Follistatin (FST) gene and its association with growth traits in native and exotic chicken.

Follistatin (FST), a member of the transforming growth factor beta super-family regulates body growth by inhibiting the binding of myostatin (an inhibitor of growth) with its receptor in chicken. An experiment was conducted to explore ontogenic expression of the follistatin gene, determine polymorphism at the coding region of the gene and estimate its effect on growth traits in native (Aseel) and exotic broiler (PD-1) and layer (White Leghorn) chicken. The significant differences of FST gene expression were observed among the breeds revealing significantly (p < 0.05) higher expression in PD-1 line followed by White Leghorn and Aseel breeds during both embryonic and post-hatch period. The polymorphism at the functional domain of the FST gene was identified with the presence of 4 haplotypes. The follistatin haplogroups had the significant effect on body weights (p < 0.05) at 42 days of age in the White Leghorn, PD-1 and Aseel breeds (h1h1 in PD-1, h1h4 in White Leghorn and h1h2 haplogroups in Aseel breeds had the highest body weights of 770.04 ± 12.96, 246.28 ± 7.60 and 270.00 ± 10.68 g, respectively). It is concluded that the follistatin gene expressed differently during the embryonic and post-embryonic period across the breeds and the coding region of the gene was polymorphic having significant effects on growth traits in chicken.

Establishment of primary chicken embryo myoblast cell culture, antigenic epitopes prediction and production of anti activin receptor type IIB polyclonal antibody in chicken.

The detection of activin receptor typeIIB (ACTRIIB) protein, a prominent negative muscle growth regulator has paramount value in augmenting growth traits through molecular breeding schemes in chicken. The study was formulated to establish primary chicken embryo myoblast culture (CEM) using 9th and 18th day chick embryos and to develop antibodies for immunodetection of ACTRIIB protein. The physicochemical and structural attributes of the ACTRIIB sequence were evaluated to identify substantial antigenic regions. The ACTRIIB sequence was transfected into CEM and expressed protein was injected subcutaneously into rats to produce hyperimmune serum. The average propensity of protein sequence for beta turns, surface accessibility, chain flexibility, antigenicity, hydrophilicity and linear epitopes was 0.978, 1.000, 0.991, 1.038, 1.258 and 0.512, respectively. The 9th day CEM exhibited confluency (80-90%) earlier than the 18th day. The expression of myogenic regulatory factors in 9th day myoblasts was higher than the 18th day by 7.28, 5.16, 6.28 and 6.93 folds for MYF5, MRF4, MYOG and MYOD, respectively. The ACTRIIB mRNA was downregulated by 2.54 folds on the 9th day compared to the 18th day myoblasts and protein varied significantly between 9th and 18th day myoblasts. The CEM culture can be harnessed unequivocally to investigate molecular mechanisms underlying muscle growth besides raising antibodies.

Cloning, characterisation and expression of the SERPINB14 gene, and association of promoter polymorphisms with egg quality traits in layer chicken.

1. Ovalbumin (SERPINB14) is the most abundant protein present in egg white contributing about 54% of the total egg protein. In this study, the objectives were to clone and characterise the coding sequence of the SERPINB14 gene, to explore its expression profile, identify polymorphisms in the promoter of the gene and explore any association with egg quality traits in White Leghorn chickens.2. SNPs and mRNA expression of SERPINB14 in White Leghorn chicken lines were detected by PCR-single strand conformation polymorphism (SSCP) along with sequencing and qPCR. The open reading frame (ORF) was cloned in an expression plasmid vector and sequenced.3. The ORF of this gene was 1161 bp encoding a peptide of 386 amino acids. There were three SNPs observed in the coding region of the gene, one of which was of the mis-sense type, having c562G>A transition which resulted in substitution of alanine to threonine at position 188 in the protein sequence. In both the lines, an increase in expression of the gene was observed after onset of egg production with peak expression at the 40th week of age compared to before onset of lay. The SERPINB14 gene was expressed in the magnum, but not in ovary and infundibulum, tissues of each White Leghorn line. The promoter region of the gene showed SNPs with three haplotypes; H1, H2, and H3. The haplo groups were associated with the egg weight and age at sexual maturity in the IWI line and Haugh unit and albumin index in the IWK line.4. It was concluded that the ORF of SERPINB14 gene in White Leghorn chicken lines is polymorphic. The promoter region of the gene is also polymorphic and has significant (P < 0.05) association with Haugh unit and egg weight in IWK and IWI chicken lines, respectively.

In silico prediction of short hairpin RNA and in vitro silencing of activin receptor type IIB in chicken embryo fibroblasts by RNA interference.

Gene silencing by RNA interference is extensively used reverse genetic approach to analyse the implications of any gene in mammalian systems. The silencing of the Activin type IIB receptor belonging to transforming growth factor beta superfamily has demonstrated increase in muscle growth in many species. We designed five short hairpin RNA constructs targeting coding region of chicken ACTRIIB. All the shRNAs were transfected into chicken embryo fibroblast cells and evaluated their silencing efficiency by real time PCR and western blotting. Initially the computational analysis of target region and shRNA constructs was undertaken to predict sequence based features (secondary structures, GC% and H-b index) and thermodynamic features (ΔGoverall, ΔGduplex, ΔGbreak-target, ΔGintra-oligomer, ΔGinter-oligomer and ΔΔGends). We determined that all these predicted features were associated with shRNA efficacy. The invitro analysis of shRNA constructs exhibited significant (P < 0.05) reduction in the levels of ACTRIIB at mRNA and protein level. The knock down efficiency of shRNAs varied significantly (P < 0.001) from 83% (shRNA 1) to 43% (shRNA 5). All the shRNAs up regulated the myogenic pathway associated genes (MyoD and MyoG) significantly (P < 0.05). There was significant (P < 0.05) up-regulation of IFNA, IFNB and MHCII transcripts. The ACTRIIB expression was inversely associated with the expression of myogenic pathway and immune response genes. The anti ACTRIIB shRNA construct 1 and 3 exhibited maximum knock down efficiency with minimal interferon response, and can be used for generating ACTRIIB knockdown chicken with higher muscle mass.

Polymorphisms in GnRHI and GnRHII genes and their association with egg production and egg quality traits in chicken.

1. Two candidate genes, namely, Gonadotropin releasing hormone I (GnRHI) and Gonadotropin releasing hormone II (GnRHII) play pivotal roles in ovulation and egg production in chicken. The objective of this study was to explore polymorphism in these genes and to estimate the effects of polymorphism of these two genes on egg production and egg quality traits in White Leghorn laying hens. 2. Single strand conformation polymorphism followed by sequencing was performed to detect polymorphism in these genes. 3. The coding regions of the GnRHI and GnRHII genes were found to be polymorphic. In the GnRH1 gene, 12 haplotypes were determined, of which the h1 haplotype was predominant and the h5, h9 and h11 haplotypes were the least frequent ones. In the GnRHII gene, eight haplotypes were found, of which the h1 haplotype was the most frequent and the h6 was the least frequent haplotype in the White Leghorn population. 4. The haplogroups of GnRHI had a significant effect on body weight and egg production up to 64 weeks of age, yolk content, Haugh units and egg shell parameters. The h1h2 haplogroup of the GnRHI gene showed the highest egg production, with 211.0 ± 24.3 eggs up to 64 weeks of age, while the highest yolk content and Haugh unit was found in h3h10 haplogrouped birds. The haplogroups of GnRHII had a significant effect on age at sexual maturity (ASM) where the shortest ASM was found in the h1h4 birds (147.3 ± 5.9 d) and the longest ASM was observed in the h1h3 birds (160.6 ± 23.4 d). 5. It was concluded that GnRHI and GnRHII genes are polymorphic and have a significant effect on body weight, egg production and egg quality traits in White Leghorn laying hens.

Co-existence of psoriasis and melanoma in a large urban academic centre population: a cross-sectional retrospective study.

BACKGROUND: Psoriasis has been linked to increased malignancy risk, particularly lympho-haematopoietic and non-melanoma skin cancers; however, its association with cutaneous melanoma remains unclear. OBJECTIVE: The aim of this study was to determine if there is an association between melanoma and psoriasis in a large, urban academic population through an electronic medical record database. METHODS: We searched our institution's electronic medical record database (EDW-Electronic Data Warehouse) from 1/2001 to 11/2013. Subjects were identified by ICD-9 codes. Melanoma diagnosis was included only if documented at least 1 month after the psoriasis diagnosis was documented. Odds ratio (OR) was obtained for association between cutaneous melanoma and psoriasis. The OR was then adjusted for phototherapy and age. To minimize detection bias, we also obtained the OR for association between cutaneous melanoma and atopic dermatitis. RESULTS: We identified 10 947 patients with psoriasis, 64 of whom had a subsequent diagnosis of cutaneous melanoma. We detected a significant association between melanoma and psoriasis (OR = 1.77; 95%CI 1.38-2.26; P < 0.0001; total n = 1 525 252). After adjusting for phototherapy and age, a statistically significant association between melanoma and psoriasis remained detectable (OR = 1.9; 95%CI 1.55-2.55; P < 0.0001 and OR = 1.64; 95%CI 1.17-2.26; P = 0.003 respectively). The OR for melanoma with atopic dermatitis in the same patient database showed a statistically significant inverse association between the two diseases (OR = 0.35; 95%CI 0.16-0.73; P = 0.005). CONCLUSION: Our findings show a statistically significant association between psoriasis and melanoma. After adjusting the OR for phototherapy and age, a statistically significant association remained. Further investigations exploring these associations are warranted in order to establish the relative risk for melanoma in psoriasis patients.

Gene Expression and Polymorphism of Myostatin Gene and its Association with Growth Traits in Chicken.

Myostatin is a member of TGF-ß super family and is directly involved in regulation of body growth through limiting muscular growth. A study was carried out in three chicken lines to identify the polymorphism in the coding region of the myostatin gene through SSCP and DNA sequencing. A total of 12 haplotypes were observed in myostatin coding region of chicken. Significant associations between haplogroups with body weight at day 1, 14, 28, and 42 days, and carcass traits at 42 days were observed across the lines. It is concluded that the coding region of myostatin gene was polymorphic, with varied levels of expression among lines and had significant effects on growth traits. The expression of MSTN gene varied during embryonic and post hatch development stage.

Gene expression and silencing of activin receptor type 2A (ACVR2A) in myoblast cells of chicken.

Activin receptor type 2A (ACVR2A) acts as receptor for myostatin (MSTN) protein involved in inhibiting satellite cell proliferation and differentiation. The importance of the ACVR2A gene during embryonic and post-hatch periods in broiler and layer chicken was studied in an in vitro cell culture system. The expression pattern of the ACVR2A gene during embryonic stages was similar in broiler and layer lines. Post-hatch expression of the ACVR2A gene varied significantly between broiler and layer lines. Five shRNA molecules were designed to knockdown expression of the ACVR2A gene in chicken myoblast cells. The silencing of the ACVR2A gene in a cell culture system varied from 60% to 82%. It is concluded that between broiler and layer lines, there were no significant changes in expression of the ACVR2A gene during embryonic stages but it varied significantly during the post-hatch period. The shRNA showed silencing of the ACVR2A gene under an in vitro cell culture system.

Cloning, characterization and expression of myostatin (growth differentiating factor-8) gene in broiler and layer chicken (Gallus gallus).

A study was conducted to characterize myostatin gene in broiler and layer chicken and to explore mRNA expression profile in these two varieties of chicken. The myostatin cDNAs of broiler and layer varieties were cloned and sequenced. The total length of the cDNA was 1128 bp. The differences of nucleotides between PB-1 broiler and IWI layer were C > 65 > T, C > 306 > T and C > 1094 > T while those between CB broiler and IWI layer were C > 65 > T, C > 195 > G, G > 234 > A, C > 306 > T, T > 939 > C, C > 961 > T, G > 966T and C > 1094 > T. The amino acid differences of myostatin protein between PB-1 and IWI strains were alanine > 22 > valine and proline > 365 > leucine while those between CB and IWI strains were alanine > 22 > valine, histidine > 321 > tyrosine and proline > 365 > leucine. The phylogenetic study revealed closeness of PB-1 and control broiler forming a cluster, which was also closely related to IWI layer chicken formed a separate cluster. The gene was cloned and expressed in E. coli. The gene expression profile in muscle was different between broiler and layer strains. Between two broiler strains, the pattern of expression was similar. Between IWI layer and either PB-1 or CB broilers, differences in expression was found at different time points, particularly at second, fourth and seventh weeks of age. The myostatin expression was significantly associated with body weights in chicken. It is concluded that myostatin gene sequences varied at nucleotide as well as amino acid level between broiler and layer chicken varieties and the gene also expressed differentially in these two varieties.

Polymorphism and expression of insulin-like growth factor 1 (IGF1) gene and its association with growth traits in chicken.

1. The objectives of the study were to detect polymorphism in the coding region of the IGF1 gene, explore the expression profile and estimate association with growth traits in indigenous and exotic chickens. 2. A total of 12 haplotypes were found in Cornish, control layer and Aseel breeds of chicken in which the h1 haplotype was most frequent. 3. Nucleotide substitutions among haplotypes were found at 21 positions in the IGF1 gene in which 4 substitutions resulted in non-synonymous mutations in the receptor binding domain of the IGF1 protein. 4. The haplogroup showed a significant effect on body weight at 24 and 42 d of age in the control layer line, body weight at 42 d and daily weight gain between 29 and 42 d in the control broiler line, daily weight gain between 29 and 42 d in Cornish, and body weights at 42 d as well as daily weight gain between 29 and 42 d in Aseel birds. 5. IGF1 expression varied among the breeds during embryonic and post-hatch periods. The expression among the haplogroups varied in different chicken tissues. The effect of haplogroup on myofibre number in pectoral muscle was non-significant, although there was significant variation in numbers between d 1 and d 42, and between broiler and layer lines. 6. It was concluded that the coding region of the IGF1 gene was polymorphic, expressed differentially during the pre-hatch and post-hatch periods, and haplogroups showed significant association with growth traits in chicken.

Milk somatic cell DNA isolation and characterization of κ-casein gene in Halari donkey milk.

Halari donkey breed is one of the indigenous breeds of India and its population is rapidly decreasing. The Jenny milk is more similar to human milk, exhibits a wide range of probiotic diversity and hypo-allergenicity, especially among infants suffering from cow and buffalo milk protein allergy. Some studies indicated low levels of κ-casein fraction of casein protein in donkey milk. The k-casein gene was not amplified from the DNA derived from the milk somatic cells of Halari donkeys. The Halari donkey milk has low somatic cells count. We report the first isolation of DNA from milk somatic cells of Halari donkeys with subsequent characterization of k-casein gene. Through our work, we showed that the milk somatic cells can be used as a non-invasive source for DNA isolation towards molecular studies. It also proved the presence of k-casein gene in Halari donkey milk by its amplification from isolated DNA.

Polymorphism of the myostatin gene and its association with growth traits in chicken.

An experiment was carried out on myostatin gene with the objectives of identification of polymorphism in the myostatin gene and estimation of the effect of polymorphism on growth traits in chickens. Single-stranded conformation polymorphism followed by sequencing was performed to reveal polymorphism of the gene. A total of 13 haplotypes were observed across 3 chicken lines (PB-1 and CB as broiler lines and IWI as the layer line). Myostatin haplogroups had a significant effect on BW at 28, 42, and 49 d of age in the PB-1 line. The significant association of haplogroups was observed with BW at d 14 and 49 in the CB line. In the IWI layer line, the myostatin gene was polymorphic but had no significant association with growth traits. It is concluded that the myostatin gene was polymorphic and had a significant effect on growth traits in broiler chickens.

Indian Red Jungle fowl reveals a genetic relationship with South East Asian Red Jungle fowl and Indian native chicken breeds as evidenced through whole mitochondrial genome sequences.

Background: Native chickens are dispersed in a wide geographical range and have hereditary assets that are kept by farmers for various purposes. Mitochondrial DNA (mtDNA) is a widely utilized marker in molecular studies because of its quick advancement, matrilineal legacy, and simple molecular structure. Method and Results: We performed NGS sequencing to investigate mitochondrial genomes and to evaluate the hereditary connections, diversity, and measure of gene stream estimation in Indian native chicken breeds and Red Jungle fowl. The chicken breeds were genotyped using the D-loop region and 23 haplotypes were identified. When compared to Indian native breeds, more haplotypes were identified in the NADH dehydrogenase subunits, Cytochrome c oxidase, Cytochrome b, ATP synthase subunit 6, and Ribosomal RNA genes. The phylogenetic examination indicated that the analyzed chicken breeds were divided into six significant clades, namely A, B, C, D, E, and F, of which the F clade indicated the domestication of chicken breeds in India. Additionally, our work affirmed that the Indian Red Jungle Fowl is the origin for both reference Red Jungle Fowl as well as all Indian breeds, which is reflected in the dendrogram as well as network analysis based on the whole mtDNA and D-loop region. Indian Red Jungle Fowl is distributed as an outgroup, suggesting that this ancestry was reciprocally monophyletic. Conclusion: The mtDNA sequences of Indian native chickens provided novel insights into adaptation mechanisms and the significance of important mtDNA variations in understanding the maternal lineages of native birds.

A note on two-sample tests for comparing intra-individual genetic sequence diversity between populations.

Gilbert, Rossini, and Shankarappa (2005, Biometrics 61, 106-117) present four U-statistic based tests to compare genetic diversity between different samples. The proposed tests improved upon previously used methods by accounting for the correlations in the data. We find, however, that the same correlations introduce an unacceptable bias in the sample estimators used for the variance and covariance of the inter-sequence genetic distances for modest sample sizes. Here, we compute unbiased estimators for these and test the resulting improvement using simulated data. We also show that, contrary to the claims in Gilbert et al., it is not always possible to apply the Welch-Satterthwaite approximate t-test, and we provide explicit formulas for the degrees of freedom to be used when, on the other hand, such approximation is indeed possible.

Polymorphisms of Pit-1 gene and its association with growth traits in chicken.

The Pit-1 gene is involved in regulation of muscle growth through controlling the expression of growth hormone, prolactin, and transforming growth factor-ß genes in chicken. The objectives of the study were to explore polymorphisms of the Pit-1 gene and to estimate the effect of these polymorphisms on growth traits in PB-1 and control (broiler strain) and IWI (layer strain) chickens. Single-stranded conformation polymorphism followed by sequencing was performed to reveal polymorphisms of the gene. In total, 10 haplotypes were found across the lines. The mRNA expression of Pit-1 varied among haplogroups and had a significant effect on BW and growth rates. The haplogroups showed a significant effect on BW in wk 7 in PB-1 chickens. In control chickens there was a significant effect at d 1 and in wk 2 and 7, and in IWI strains, there was a significant effect at d 1 and wk 6 and 7. The significant association of haplogroups and growth rate was found between 0 and 2 wk in control and between 0 and 2 and 6 and 7 wk in IWI strains. It was concluded that the Pit-1 gene is polymorphic and has a significant effect on growth traits in chickens.

Embryonic transcriptome unravels mechanisms and pathways underlying embryonic development with respect to muscle growth, egg production, and plumage formation in native and broiler chickens.

Background: Muscle development, egg production, and plumage colors are different between native and broiler chickens. The study was designed to investigate why improved Aseel (PD4) is colorful, stronger, and grew slowly compared with the control broiler (CB). Methods: A microarray was conducted using the 7th-day embryo (7EB) and 18th-day thigh muscle (18TM) of improved Aseel and broiler, respectively. Also, we have selected 24 Gallus gallus candidate reference genes from NCBI, and total RNA was isolated from the broiler, improved Aseel embryo tissues, and their expression profiles were studied by real-time quantitative PCR (qPCR). Furthermore, microarray data were validated with qPCR using improved Aseel and broiler embryo tissues. Results: In the differential transcripts screening, all the transcripts obtained by microarray of slow and fast growth groups were screened by fold change ≥ 1 and false discovery rate (FDR) ≤ 0.05. In total, 8,069 transcripts were differentially expressed between the 7EB and 18TM of PD4 compared to the CB. A further analysis showed that a high number of transcripts are differentially regulated in the 7EB of PD4 (6,896) and fewer transcripts are differentially regulated (1,173) in the 18TM of PD4 compared to the CB. On the 7th- and 18th-day PD4 embryos, 3,890, 3,006, 745, and 428 transcripts were up- and downregulated, respectively. The commonly up- and downregulated transcripts are 91 and 44 between the 7th- and 18th-day of embryos. In addition, the best housekeeping gene was identified. Furthermore, we validated the differentially expressed genes (DEGs) related to muscle growth, myostatin signaling and development, and fatty acid metabolism genes in PD4 and CB embryo tissues by qPCR, and the results correlated with microarray expression data. Conclusion: Our study identified DEGs that regulate the myostatin signaling and differentiation pathway; glycolysis and gluconeogenesis; fatty acid metabolism; Jak-STAT, mTOR, and TGF-ß signaling pathways; tryptophan metabolism; and PI3K-Akt signaling pathways in PD4. The results revealed that the gene expression architecture is present in the improved Aseel exhibiting embryo growth that will help improve muscle development, differentiation, egg production, protein synthesis, and plumage formation in PD4 native chickens. Our findings may be used as a model for improving the growth in Aseel as well as optimizing the growth in the broiler.

Silencing acetyl-CoA carboxylase A and sterol regulatory element-binding protein 1 genes through RNAi reduce serum and egg cholesterol in chicken.

Cholesterol is synthesized in chicken through de novo lipid biosynthetic pathway where two most important genes viz. SREBP1 and ACACA play immense role. To minimize cholesterol synthesis, RNAi approach was adopted and accordingly, we developed transgenic chicken possessing ACACA and SREBP1 shRNA constructs, which showed lower level of ACACA and SREBP1 in serum. The serum total cholesterol, triglycerides, HDL and LDL cholesterol was significantly lower by 23.8, 35.6, 26.6 and 20.9%, respectively in SREBP1 transgenic birds compared to the control. The egg total cholesterol and LDL cholesterol content was numerically lower in both ACACA and SREBP1 transgenic birds by 14.3 and 13.2%, and 10.4 and 13.7%, respectively compared to the control. It is concluded that the protocol was perfected to develop transgenic chicken through RNAi for knocking down the expression of ACACA and SREBP1 proteins, which minimized the cholesterol and triglycerides contents in serum and eggs.

Polymorphism in the prolactin promoter and its association with growth traits in chickens.

The pituitary hormone prolactin has a wide variety of functions involving growth, behavioral, and ovarian activities in chickens. The objectives of the present study were to identify polymorphisms in the prolactin promoter and estimate their effects on growth traits in White Leghorn chickens. Among 28 haplotypes found, the h1 haplotype was predominant. Body weight at 16 and 64 weeks and age at sexual maturity were significantly associated with haplotype combinations (P < 0.05). The h1/h1 haplogroup showed the highest body weight at 16 weeks of age, and h1/h7 was the highest at 64 weeks. The lowest age at sexual maturity was found in birds with the h1/h6 haplotype combination, and mRNA expression of prolactin was lowest in h1/h4 birds and highest in h1/h5 birds. The prolactin promoter was highly polymorphic and had significant associations with growth traits in White Leghorn chickens.

Identification of haplotypes in promoter of prolactin gene and their effect on egg production and quality traits in layer chicken.

Expression of prolactin hormone is a crucial event in regulating egg production in chickens for which promoter plays the vital role in expressing the prolactin gene. The objective of the present study was to identify haplotypes in the prolactin promoter and their effects on egg production and egg quality traits in White Leghorn chicken. Single stranded conformation polymorphism followed by sequencing was conducted to explore polymorphism at 561 bp promoter of prolactin gene. The effect of haplotype combinations on egg production and quality traits were estimated following general linear model technique. The expression of prolactin by different haplogroups was quantified by qPCR. Total 28 haplotypes were found in White Leghorn chicken of which h1 haplotype possessed the highest frequency of 0.46 and h8, h14, h16, h25, h26, and h28 haplotypes had the lowest frequency (0.1%). The egg production up to 52 and 64 weeks of age were found to be significantly (p < 0.05) associated with haplotype combinations where the highest 52-w (52 weeks) egg production was found in animals with h1/h22 combination and the lowest production was observed in the birds with h1/h2 haplogroup. The haplotype combinations had the significant effect (p < 0.05) on Haugh Unit, yolk index and albumen weight at 40 weeks of age; Haugh Unit and albumen weight at 52 weeks of age and Haugh unit, yolk weight and yolk percentage at 64 weeks of age. The prolactin expression in h1/h22 birds was found to be the lowest and in h1/h5 birds to be the highest. The prolactin expression showed significant effect on 52-w egg production and albumin weight at 52 weeks age. In conclusion, it may be stated that the prolactin promoter was highly polymorphic and had the significant association with egg production and quality traits in White Leghorn chicken.

Association of polymorphism in the prolactin promoter and egg quality traits in laying hens.

Single strand conformation polymorphism analysis and DNA sequencing was performed in White Leghorn hens to explore the polymorphisms present in the promoter of the prolactin gene. The effects of different genotypes on egg production and quality traits were determined, and expression of the prolactin gene in different genotypes was quantified by real time-PCR. Five genotypes and four alleles at each of two Fragments of the promoter were found, of which the FG genotype in Fragment 1 and the PQ genotype in Fragment 2 were the most predominant genotypes. The genotypes of Fragment 1 had significant effects (P < 0·05) on Haugh unit, albumen weight, albumen percentage and shell percentage at 40 weeks of age; egg weight and yolk index at 52 weeks of age; and egg weight at 64 weeks of age. Prolactin expression in the genotypes of Fragment 1 differed significantly and GH genotyped birds had the highest level of expression. The genotypes of Fragment 2 did not show any significant differences of expression. It was concluded that the prolactin gene promoter was highly polymorphic, and had significant effects on egg quality traits in White Leghorn hens.