[Detection and application of PIS genetic deficiency gene in dairy goat].

The purpose of this study was to develop a molecular method for detecting polled intersex syndrome (PIS) genetic deficiency gene in dairy goat. Three pairs of primers, PIS-, PIS+, and NEI were designed based on PIS gene sequence (AF404302) to identify the PIS genetic deficiency genotype. For the normal phenotype, the fragments of 141 and 300 bp were obtained for the genotype PIS-PIS-, and 141, 449, and 300 bp for the genotype PIS-PIS+. For the PIS goat with the genotype PIS+PIS+, 449 and 300 bp were obtained. Two hundred and twenty-four dairy goats in one population were tested based on this method. The results showed that there were 150 PIS-PIS+, 70 PIS -PIS-, and 4 PIS+PIS+. The genotype frequency of PIS-PIS+ was 66.9%, and the gene frequency of PIS+ was 35.3% in the population. Therefore, the frequency of PIS offspring was over 12%. This study developed a method to detect PIS genetic deficiency dairy goat. The method could identify buck genotype accurately to avoid the occurrence of PIS genetic deficiency. The ease and accuracy show a strong potential of the method for use in marker assisted selection of dairy goats and healthy development of dairy goat industry.

[A method for screening microRNA target genes in goat skin by microRNA seed sequence mediated PCR].

The purpose of the present study was to establish a new microRNA seed mediated controllable genetic operation, namely MicroRNA Targets Finger Print (MTFP), for screening microRNA targets and detecting target gene expression profiles. Based on combining the complementary sequence of seed sequence, upstream and downstream anchor sequence including special adaptor, microRNA targets were amplified by means of the reverse transcription and special two step PCR. The polyacrylamide gel electrophoresis was used to analyze the sizes of amplified microRNA targets and their abundance of expression, which were used to screen specifically expressed genes in different physiological or experimental conditions. Specific target genes were obtained through isolation of DNA fragments and sequencing methods. As an example, by screening the targets of miR-203 in goat skin, five genes were amplified and sequenced with the sizes of 718 bp (JN709494), 349 bp (JN709495), 243 bp (JN709496), 159 bp (JN709497), and 97 bp (JN709498) from goat skin collections. The novel universal MTFP method could be applied for finding microRNA regulation targets or assessing target gene expression profile.