RESUMEN
Analysing pig class II mayor histocompatibility complex (MHC) molecules is mainly related to antigen presentation. Identifying frequently-occurring alleles in pig populations is an important aspect to be considered when developing peptide-based vaccines. Colombian creole pig populations have had to adapt to local conditions since entering Colombia; a recent census has shown low amounts of pigs which is why they are considered protected by the Colombian government. Commercial hybrids are more attractive regarding production. This research has been aimed at describing the allele distribution of Colombian pigs from diverse genetic backgrounds and comparing Colombian SLA-DRB1 locus diversity to that of internationally reported populations. Twenty SLA-DRB1 alleles were identified in the six populations analysed here using sequence-based typing. The amount of alleles ranged from six (Manta and Casco Mula) to nine (San Pedreño). Only one allele (01:02) having > 5% frequency was shared by all three commercial line populations. Allele 02:01:01 was shared by five populations (around > 5% frequency). Global FST indicated that pig populations were clearly structured, as 20.6% of total allele frequency variation was explained by differences between populations (FST = 0.206). This study's results confirmed that the greatest diversity occurred in wild boars, thereby contrasting with low diversity in domestic pig populations.
Asunto(s)
Variación Genética , Genética de Población , Antígenos de Histocompatibilidad Clase II/genética , Alelos , Animales , Cruzamiento , Colombia , Frecuencia de los Genes , Haplotipos , Filogenia , Filogeografía , Sus scrofa/genética , PorcinosRESUMEN
To improve the stability and recyclability of enzymes immobilized on metal-organic frameworks (MOFs), graphene oxide (GO) with surface oxygen-rich functional groups was selected to form ZIF-8/GO nanocomposites with the zeolitic imidazolate framework (ZIF-8) for cytochrome c (Cyt c) immobilization. It was found that the functional groups on the GO surface were involved in the growth of ZIF-8 without affecting the crystal structure but their particle size was reduced to about 200 nm. The storage stability and resistance to organic solvents of Cyt c were obviously improved after the immobilization on the ZIF-8/GO nanocomposite. On one hand, compared with Cyt c@ZIF-8 and Cyt c@GO with 30 and 60% protein leakage, Cyt c@ZIF-8/GO displayed little protein leakage after 60 h of storage. On the other hand, Cyt c@ZIF-8/GO retained a residual activity of approximately 100% after being stored in ethanol and acetone for 2 h, whereas the free enzyme, Cyt c@ZIF-8, and Cyt c@GO retained only about 10, 50, and 40%, respectively. In addition, the Cyt c@ZIF-8/GO nanocomposites can be utilized up to four cycles with virtually no loss of activity and may be further applied on H2O2 biosensing systems. The synergistic effect between MOFs and GO in ZIF-8/GO nanocomposites provides infinite possibilities as immobilized enzyme carriers.
RESUMEN
The development of an orientation immobilization technique via affinity between polyhistidine tags and metal ions aims at maintaining biocatalytic activity of the enzymes. In this work, to tackle the issue of the immobilization of adenylate cyclase (AC), a simple and effective approach of synthesizing iminodiacetic acid (IDA)-Ni2+ particles was applied for simultaneously purifying and immobilizing his-tagged AC. We chose agarose particles as carriers, and then decorated them with IDA, leading to the formation of a coordination combination of Ni2+. The porous carriers with a large pore size of 50â¯nm and a specific surface area of 45.8â¯m2/g exhibited favorable enzymatic activity and loading capacity. The optimal pH of the immobilized enzyme increased from 8.0 to 9.0 and the optimal temperature increased from 30⯰C to 35⯰C, compared to the free AC. Moreover, the immobilized AC retained a residual activity of approximately 80% after storing it at 25⯰C for 48â¯h, whereas only 40% of the activity was left in the free AC at the same conditions. Maximum yield of cyclic adenosine-3', 5'- monophosphate (cAMP) reached up to the summit of the reaction. The immobilized AC by affinity adsorption will provide a promising route for the industrial production of cAMP.