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家蚕BmUFP未知基因的序列分析和原核表达

发表时间:2010-10-15  浏览量:1840  下载量:636
全部作者: 胡波,张耀洲
作者单位: 浙江理工大学生物化学研究所;浙江省家蚕生物反应器和生物医药重点实验室
摘 要: 从本实验室构建的家蚕cDNA文库中克隆到一条新的基因序列,GenBank登录号为BAD05929,经Blastn及氨基酸序列同源性比对后,发现该基因是一个未知功能序列,并命名为BmUFP.该基因全长774 bp,由516 bp的开放阅读框序列、67 bp的 5′端非翻译区序列和191 bp的3′端非翻译区序列组成,开放阅读框(open reading frame,ORF)共编码171个氨基酸,无内含子。编码的BmUFP蛋白预测分子量为18.49 ku,等电点为8.72,是一个碱性蛋白,且含有一个Reeler Superfamily保守结构域。将BmUFP基因片段克隆到原核表达载体pET28a中,构建重组载体,并转化大肠杆菌Rosetta(DE3),经异丙基-β-D-硫代半乳糖苷(IPTG)诱导后,成功表达了BmUFP蛋白,为进一步研究此基因的功能奠定了基础。
关 键 词: 分子生物学;BmUFP蛋白;原核表达;家蚕;新基因
Title: Sequence analysis and prokaryotic expression of a novel BmUFP gene from silkworm, Bombyx mori
Author: HU Bo, ZHANG Yaozhou
Organization: Institute of Biochemistry, Zhejiang Sci-Tech University; Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine
Abstract: A novel gene sequence was obtained from silkworm pupa cDNA library constructed by our laboratory, and the GenBank accession number was BAD05929. After Blastn based on NCBI database and alignment of deduced amino acid sequence homology, it was found to be an unknown gene, and named as BmUFP. The full length of the gene was 774 bp. It contained an open reading frame (ORF) of 516 bp, 67 bp nucleotide sequence in 5′untranslated region (UTR) and 191 bp nucleotide sequence in 3′UTR, and no introns. The BmUFP gene encoded 171 amino acids, with their predicted mass 18.49 ku and isoelectric point 8.72, a basic protein. It was predicted that BmUFP contained a putative conserved domain of Reeler Superfamily. The ORF fragment of BmUFP gene and pET28a expression vector were ligated. The soluble BmUFP protein induced by IPTG in E. coli Rosetta (DE3) was expressed, which may lay a foundation for the study of BmUFP gene function.
Key words: molecular biology; BmUFP protein; prokaryotic expression; silkworm; novel gene
发表期数: 2010年10月第19期
引用格式: 胡波,张耀洲. 家蚕BmUFP未知基因的序列分析和原核表达[J]. 中国科技论文在线精品论文,2010,3(19):1966-1973.
 
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