水牛HSD17B1基因克隆分析及其真核表达载体构建

doi:10.16431/j.cnki.1671-7236.2016.03.001

Abstract

Abstract: In order to clarify the effect of 17 beta-hydroxysteroid dehydrogenase 1 (HSD17B1) gene on the reproductive performance of buffalo,in this study,buffalo HSD17B1 gene was cloned by 3'-RACE,analyzed by bioinformatics,and studied with eukaryotic vector construction and cell transfection technology.The results showed that the coding region of buffalo HSD17B1 was 954 bp,3'-UTR was 58 bp,encoded 317 amino acids.The buffalo HSD17B1 gene shared 100%,100%,92%,94%,87%,87% and 87% of similar nucleotide sequence with that of Bos taurus,Ovis aries,Sus scrofa,Equusca ballus,Canis lupus,Loxodonta africana and Homo sapiens,respectively.Phylogenetic tree analysis showed that HSD17B1 gene was highly conserved in different species and evolution.HSD17B1 protein was weakly acidic,without signal peptide,located inthe cytoplasmic,and with the presence of type1_17beta-HSD-like_SDR_c,PRK05993,LPOR and FabG domains.The buffalo HSD17B1 eukaryotic expression vector was successfully constructed,after transfected into 293T cell lines,HSD17B1-EGFP fusion protein was detectable.In a word,the success cloning and construction of eukaryotic expression vector of buffalo HSD17B1 gene,provided an important reference for surveying the regulation mechanism of HSD17B1 gene during buffalo folliculogenesis and embryogenesis.

Key words: buffalo; HSD17B1 gene; cloning and bioinformatics analysis; construction of vector

CLC Number:

ZHU Peng, PANG Chun-ying, DENG Ting-xian, DUAN An-qin, LU Xing-rong, CHEN Ming-tang, YANG Bing-zhuang, LIANG Xian-wei. Cloning and Construction of Eukaryotic Expression Vector of Buffalo HSD17B1 Gene[J]. , 2016, 43(3): 559-567.

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Cloning and Construction of Eukaryotic Expression Vector of Buffalo HSD17B1 Gene

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