灰飞虱actin基因的克隆及原核表达  

Cloning and prokaryotic expression of actin gene in Laodelphax striatellus Fallén

作　　者：陈晴晴[1,2] 李艳舞 倪海平[2] 徐秋芳[2] 张金凤[2] 李硕[2] 周益军[2] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210046 [2]江苏省农业科学院植物保护研究所,江苏南京210014

出　　处：《江苏农业学报》2014年第5期1010-1014,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31170142;31471768);公益性行业(农业)科研专项(201003031);江苏省农业科技自主创新基金项目[CX(13)5019]

摘　　要：通过生物信息学分析获得了灰飞虱actin基因的序列信息,采用RT-PCR方法从灰飞虱中克隆了actin基因的开放阅读框,并将其在大肠杆菌BL21(DE3)中进行原核表达。序列分析结果表明,该基因开放阅读框长1 131 bp,编码377个氨基酸,蛋白质理论分子量为4.17×104,理论等电点为5.28。将此序列登录GenBank数据库,登录号为KC683802。序列比对发现,该基因序列在多个物种间保守,与褐飞虱actin基因的同源性高达95%。构建的原核表达载体pET32a-actin,在大肠杆菌BL21(DE3)中进行获得表达,SDS-PAGE分析结果表明,表达的蛋白质主要存在于包涵体中。To study the role of actin gene during the interaction between small brown planthopper （ SBPH, Laodel-phax striatellus Fall＆#233;n） and rice black- streaked dwarf virus （RBSDV）, the sequence information of actin gene was ob-tained by bioinformatics analysis. The complete open reading frame （ ORF） was cloned from SBPH by reverse transcription polymerase chain reaction （RT-PCR） and was expressed in Escherichia coli BL21（DE3）. Sequence analysis showed that actin ORF is 1 131 base pairs （ bp） in length encoding a polypeptide of 377 amino acids with the theoretical molecular weight of 4. 17＆#215; 104 and the theoretical pI of 5. 28. The sequence was submitted to GenBank and accession number is KC683802. Sequence alignment indicated that actin is conserved among several species. The sequences of SBPH actin shared 95% similarity with Nilaparvata lugens actin. The SDS-PAGE results showed that the actin fusion protein was ex-pressed in the form of inclusion body.

关 键 词：灰飞虱 肌动蛋白 基因克隆 原核表达 

分 类 号：S433.3[农业科学—农业昆虫与害虫防治]