版纳微型猪近交系DLDH基因编码区序列扩增及生物信息学分析

Amplification and Bioinformatics Analysis of Coding Region of the DLDH Gene in Banna Mini-pig Inbred Line(BMI)

本试验旨在扩增版纳微型猪近交系DLDH基因编码区序列,为进一步研究DLDH基因的表达信号通路及与精子获能和顶体反应发生相关基因的表达调控提供必要的基础。通过提取版纳微型猪近交系睾丸组织RNA,经反转录后获得cDNA,设计特异性引物以扩增DLDH基因的编码区序列,经测序后采用生物信息学软件对编码区序列及氨基酸序列进行生物信息学分析和结构预测。结果表明,版纳微型猪近交系DLDH基因编码区序列全长1 530bp,编码509个氨基酸。生物信息学分析表明,不同物种DLDH基因编码区核苷酸序列和氨基酸序列具有较强的保守性,DLDH蛋白中存在丰富的磷酸化位点,空间结构以α-螺旋和无规则卷曲为主。

The study aimed to amplify the coding region of DLDH gene,which could be used to further study the signal pathway of DLDH expression and the correlation between DLDH gene expression and sperm capacitation,acrosome reaction.RNA was extracted from testicle and cDNA was obtained by reverse transcription.Specific primers were designed to amplify the coding region of DLDH gene.The sequence of coding region and amino acid of DLDH were analyzed and predicted by Lasergene,BioEdit,ClustalX,Mega bio-softs,and the molecular phylogenetic trees of coding amino acids region of DLDH gene was constructed by NJ and ME,respectively.The sequence length of coding region of the BMI DLDH gene was 1 530 bp,encoding aprotein of 509 amino acids.By biology software analysis,the nucleotide and amino acid sequences of DLDH among different species were conserved in evolution.The DLDH protein contained the abundant phosphorylated sites and was assembled withα-helixes and random coil.