牛HSL基因的克隆和生物信息学分析

Cloning and Bioinformatical Analysis of the HSL Gene in Cattle

旨在克隆牛HSL基因的CDS序列,并对该基因进行生物信息学、组织表达谱分析。根据GenBank登录的牛HSL基因序列(NM_001080220)设计1对引物,对牛组织样品中的HSL基因CDS序列进行RT-PCR扩增及序列测定;利用半定量RT-PCR方法检测HSL基因在牛各组织中的表达情况;利用生物信息学软件对其所编码的牛HSL蛋白进行分析。结果显示,获得的牛HSL基因CDS全长为2271bp,编码756个氨基酸,与GenBank登录的牛HSL基因同源性最高,为99.9%。HSL蛋白含有一个HSL-Nsuperfamily保守结构域,无信号肽结构,具有疏水性。半定量RT-PCR显示,HSL基因在检测的心脏、脾脏、肺脏、肾脏、肝脏、大网膜、皮下脂肪、肌肉8种组织中均有表达,其中在大网膜和皮下脂肪中表达量较高,在肾脏、脾脏、肝脏、肺脏、肌肉和心脏中度表达。该试验可为研究牛HSL蛋白的结构和功能提供参考。

In this study, the CDS sequence of the bovine HSL gene was cloned and bioinformatically analyzed, and its tissue expression profile was characterized. One set of primers was designed based on the bovine HSL gene sequence (NM_001080220) registered in the GenBank, and the CDS sequence of HSL gene in bovine tissue samples was amplified using RT-PCR assay and was determined by sequencing;the expression level of HSL gene in bovine tissues was detected by semi-quantitative RT-PCR;the HSL protein encoded by HSL gene was bioinformatically analyzed. The results showed that the obtained CDS sequence of bovine HSL gene was 2 271 bp in length encoding 756 amino acids, and it shared the highest nucleotide homology (99.9%) with the bovine HSL gene registered in the GenBank;the HSL protein was hydrophobic and contained a HSL-N superfamily conserved domain with no signal peptide structure;semi-quantitative RT-PCR revealed that the HSL gene was expressed in 8 tissues including heart, spleen, lung, kidney, liver, greater omentum, subcutaneous fat and muscle;the highest expression level was observed in the greater omentum and subcutaneous fat, and the moderate expression level was found in kidney, spleen, liver, lung, muscle and heart. This study may provide a reference for assessing the structure and function of bovine HSL proteins.