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为研究贵州黑山羊、贵州白山羊MHC-DRB1基因遗传机制,试验运用混合DNA池结合PCR产物直接测序方法,对MHC-DRB1基因第三外显子区域进行多态性分析,利用生物信息学分析软件对PCR扩增所获序列进行m RNA二级结构及蛋白质的二级结构和抗原表位分析。经序列对比发现9个SNPs位点,其中A~(8858)G(ILe-Val)、G~(8969)A(Gly-Ser)、G~(8978)A(Glu-Lys)、T~(9094)G(Asp-Glu)4个单核苷酸突变,导致所编码氨基酸发生改变,其它5个位点均属于同义突变。进一步分析发现,A~(8858)G、C~(8974)T、T~(9094)G、C~(9100)T、G~(9124)A突变位点导致m RNA二级结构的变化,A~(8858)G以及T~(9094)G对蛋白质二级结构影响最大。
In order to study the genetic mechanism of MHC-DRB1 gene in Guizhou black goat and Guizhou white goat, the polymorphism of the third exon of MHC-DRB1 gene was analyzed by hybrid DNA pool combined with direct sequencing of PCR products. Bioinformatics analysis The software amplifies the secondary structure of m RNA and the secondary structure of protein and epitope analysis of the sequence obtained by PCR amplification. Nine SNPs were found by sequence comparison. Among them, A ~ (8858) G (ILe-Val), G8969A (Gly-Ser), G8978A (Glu-Lys) 9094) G (Asp-Glu) 4 single nucleotide mutations, resulting in the encoded amino acid changes, the other five sites are synonymous mutations. Further analysis showed that the secondary structure of m RNA was induced by A ~ (8858) G, C ~ (8974) T, T ~ (9094) G, C ~ (9100) T and G ~ (9124) The effects of A ~ (8858) G and T ~ (9094) G on protein secondary structure were the most significant.