Wang Guosheng, Zhao Xingbo, Zhong Juming, Cao Meng, He Qinghua, Liu Zhengxin, Lin Yaqiu, Xu Yaou, Zheng Yucai
College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China.
Int J Mol Sci. 2013 Jun 5;14(6):11994-2003. doi: 10.3390/ijms140611994.
The main objective of this work was to study the unique polymorphisms of the lactate dehydrogenase-1 (LDH1) gene in yak (Bos grunniens). Native polyacrylamide gel electrophoresis revealed three phenotypes of LDH1 (a tetramer of H subunit) in yak heart and longissimus muscle extracts. The corresponding gene, ldhb, encoding H subunits of three LDH1 phenotypes was obtained by RT-PCR. A total of six nucleotide differences were detected in yak ldhb compared with that of cattle, of which five mutations cause amino acid substitutions. Sequence analysis shows that the G896A and C689A, mutations of ldhb gene, result in alterations of differently charged amino acids, and create the three phenotypes (F, M, and S) of yak LDH1. Molecular modeling of the H subunit of LDH indicates that the substituted amino acids are not located within NAD+ or substrate binding sites. PCR-RFLP examination of G896A mutation demonstrated that most LDH1-F samples are actually heterozygote at this site. These results help to elucidate the molecular basis and genetic characteristic of the three unique LDH1 phenotypes in yak.
本研究的主要目的是探究牦牛（Bos grunniens）乳酸脱氢酶-1（LDH1）基因的独特多态性。非变性聚丙烯酰胺凝胶电泳显示，牦牛心脏和背最长肌提取物中存在三种LDH1表型（H亚基的四聚体）。通过RT-PCR获得了编码三种LDH1表型H亚基的相应基因ldhb。与牛的ldhb相比，牦牛ldhb共检测到6个核苷酸差异，其中5个突变导致氨基酸替换。序列分析表明，ldhb基因的G896A和C689A突变导致带不同电荷的氨基酸发生改变，从而产生了牦牛LDH1的三种表型（F、M和S）。LDH H亚基的分子建模表明，被替换的氨基酸不在NAD+或底物结合位点内。对G896A突变的PCR-RFLP检测表明，大多数LDH1-F样本在该位点实际上是杂合子。这些结果有助于阐明牦牛三种独特LDH1表型的分子基础和遗传特征。
