水牛Smad4基因的克隆及其真核表达载体的构建

doi:10.16431/j.cnki.1671-7236.2017.08.022

Abstract

Abstract:

This study was aimed to elucidate the molecular mecbanisms of Smad4 gene on granulose cells proliferation and embryonic development in buffalo. The Smad4 gene was amplified by RT-PCR, analyzed by bioinformatics, studied with eukaryotic vector construction, and used liposome transfection skill to transfect into the buffalo granulose cells. The results showed that Smad4 gene was cloned, the coding region was 1 662 bp, and encoded 553 amino acids. BLAST analysis showed that the buffalo Smad4 gene shared 99% of similar nucleotide sequence with Bos taurus, and shared 98%, 96%, 96% and 95% of similar nucleotide sequence with Ovis aries, Sus scrofa, Equues caballus and Homo sapiens, respectively. Phylogenetic tree analysis showed that Smad4 gene was highly conserved in different species. The buffalo pEGFP-N1-Smad4 eukaryotic expression vector was successfully constructed, and transferred into the buffalo granulose cells, and the Smad4-EGFP fusion protein was detected in the cells. The results suggested that the success cloning and construction vector of buffalo Smad4 gene laid the foundation for the research of Smad4 gene on embryo development.

Key words: buffalo; Smad4 gene; cloning; eukaryotic expression vector

CLC Number:

Q785

YAN Sheng-fei, ZHANG Xiu-fang, HUANG Yue-meng, ZHENG Hai-ying, YANG Chun-yan, QIN Guang-sheng, HUANG Jia-xiang, SHANG Jiang-hua. Cloning and Construction of Eukaryotic Expression Vector of Buffalo Smad4 Gene[J]. , 2017, 44(8): 2369-2377.

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Cloning and Construction of Eukaryotic Expression Vector of Buffalo Smad4 Gene

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