CAJ | 학술논문

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1131 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。
통과생물신식학분석획득료회비슬actin기인적서렬신식,채용RT-PCR방법종회비슬중극륭료actin기인적개방열독광,병장기재대장간균BL21( DE3)중진행원핵표체。서렬분석결과표명,해기인개방열독광장1131 bp,편마377개안기산,단백질이론분자량위4.17×104,이론등전점위5.28。장차서렬등록GenBank수거고,등록호위KC683802。서렬비대발현,해기인서렬재다개물충간보수,여갈비슬actin기인적동원성고체95%。구건적원핵표체재체pET32a-actin,재대장간균BL21(DE3)중진행획득표체,SDS-PAGE분석결과표명,표체적단백질주요존재우포함체중。
To study the role of actin gene during the interaction between small brown planthopper ( SBPH, Laodel-phax striatellus Fallén) 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× 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.