中华大蟾蜍γ-肌动蛋白基因Bbgγ-Actin的克隆与生物信息学分析  

Cloning and Bioinformatic Analysis of γ-Actin Gene(Bbgγ-Actin) from Bufo bufo gargarizans

作　　者：刘彦芳[1,2] 王秋爽[1,2] 凌鸿[1,2] 刘东春[1,2] 代英辉[1] 王东[1,2] 

机构地区：[1]沈阳药科大学中药学院,沈阳110016 [2]沈阳药科大学中韩分子生药学研究室,沈阳110016

出　　处：《生物技术通报》2016年第9期156-164,共9页Biotechnology Bulletin

基　　金：辽宁省自然科学基金项目(2015020741);沈阳药科大学归国人员科研启动基金项目(GGJJ2015101)

摘　　要：旨在获得中华大蟾蜍肌动蛋白序列全长。以中华大蟾蜍(Bufo bufo gargarizans)耳后腺为材料,提取总RNA,通过转录组高通量测序快速得到中华大蟾蜍肌动蛋白基因(Bbg Actin)cDNA全长2 055 bp,通过RT-PCR技术对其ORF序列进行验证,并对Bbg Actin基因进行分析。序列分析表明,Bbg Actin基因开放阅读框长1 131 bp,编码376个氨基酸。通过BLAST P程序分析,所得序列与Gen Bank数据库中肌动蛋白基因序列相似度均在89%以上,氨基酸序列的相似性达90%以上。系统进化分析表明,Bbg Actin与γ-Actin聚为一类,所得中华大蟾蜍肌动蛋白属于γ-Actin。首次获得了中华大蟾蜍γ-Actin cDNA全长序列。In order to obtain the full-length of actin＇ s sequence in Bufo bufo gargarizans, the total RNA was extracted from the parotid gland of B. bufo gargarizans, and high-throughput sequencing of transcriptome was used to obtain the full-length cDNA of actin （ BbgActin ） in B. gargarizansas 2 055 bp. The sequence of ORF （ open reading frame） was verified by RT-PCR and sequencing. Sequence analysis showed that the ORF was 1 131 bp encoding for a polypeptide of 376 amino acid residues. Homologous alignment by software BLAST P revealed that it shared over 89% of nucleotide identities and over 90% of amino acid identities with actins from other animals in GenBank. Phylogenetic analysis showed that BbgActin clustered in a class with γ-Actin and highly conserved during evolution. In conclusion, it is the first time to acquire the full-length sequence of γ-Actin cDNA in B. bufo gargarizans, which lays a foundation for further studies on the role of gene Bbgγ-Actin on the life activities of B. bufo gargarizans.

关 键 词：中华大蟾蜍 γ-肌动蛋白 CDNA全长 系统进化 

分 类 号：S865.9[农业科学—野生动物驯养]