摘要
为了克隆牛TNS1基因序列,试验以人TNS1基因序列为探针,通过NCBI在线网站BLAST获得同源性较高的牛表达序列标签(ESTs)及部分mRNA序列,采用RT-PCR方法从牛肌肉组织克隆cDNA序列并与牛ESTs和mRNA序列进行拼接,获得牛TNS1基因序列。结果表明:克隆获得的牛TNS1基因含有32个外显子和31个内含子,编码区长度为5 148 bp,编码1 715个氨基酸。该蛋白含有149个磷酸化位点,分别位于丝氨酸(Ser)、酪氨酸(Tyr)和苏氨酸(Thr)残基上,二级结构以无规则卷曲为主,含有PTEN_C2、Src同源结构域2(SH2)和磷酸化酪氨酸结合区域(PTB)3个功能结构域。牛TNS1蛋白线粒体靶向肽及分泌通路信号肽所占的比例很小,大部分分布于微体和细胞核内,不属于分泌蛋白。
In this study high- homology expressed sequence tags（ESTs） and partial mRNA sequences of bovine TNS1 gene were obtained by BLAST in NCBI GenBank according to the standard human TNS1 gene sequence. The eDNA sequence of bovine TNS1 gene which was ampli- fied by RT - PCR based on mRNA extraction of musele tissue was in assembly with the known ESTs and mRNA sequences, and the complete se- quence of bovine TNS1 gene was obtained. The results showed that the gene contained 32 exous and 31 intrens, with CDS region of 5 148 hp en- coding 1 715 amino acids. Protein analysis suggested that the bovine TNS1 gene contained 149 phosphorylation sites located at residues serine （Set） , tyrosine （Tyr）and threonine （Thr） , and also had random coils in its secondary structure, and three functional domains （PTEN C2, SH2 and PTB）. The subcellular localization analysis indicated that bovine TNS1 protein was in a very small proportion to be a mitochondrion -target peptide or a secretory pathway signal peptide,while was predominantly located in the microbedy and the nucleus,which didnt belong to the secreted protein.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2017年第8期98-102,290,共6页
Heilongjiang Animal Science And veterinary Medicine
基金
国家转基因重大专项项目(2014ZX08007-002)
