摘要
以来自湖北省的HB柚柑橘衰退病毒(Citrus tristeza virus,CTV)分离物(CTV-HB1)为材料,利用RT-PCR技术对其CP基因进行克隆,序列分析结果表明:CTV-HB1与典型茎陷点分离物SY568和NUagA的核苷酸和推定氨基酸的序列相似性均在97%以上,而与CTV速衰分离物T36和弱毒分离物T385和T30的核苷酸和氨基酸序列相似性较低,均在95%以下。将CP基因片段连接到表达载体,转化大肠杆菌后诱导重组蛋白的表达。SDS-PAGE和Westen-blot分析结果表明,经IPTG诱导后产生了预期大小的重组蛋白,该蛋白可与CTV-L5提纯病毒制备的多克隆抗体发生特异性免疫反应。
In this study, RT- PCR protocol was used for cloning the CP gene of CTV isolate from HB pummelos （ Citrus grandis） in Hubei province. Comparison of the CP gene sequence from CTV - HB1 with those of other isolates published in GenBank revealed that CTV - HB1 was closely related to stem pitting isolates SY568 and NUagA at nucleotide and deduced amino acid levels, the similarities were higher than 97 %. However, the similarities were lower than 95% between CTV - HB1 and severe isolate T36, mild isolates T30 and T385. The recombinant plasmid containing CP was transformed into E. coli BL21 （ DE3 ） plysS. Results of SDS - PAGE and Western - blot showed that the specific fusion protein was induced to express by IPTG, in addition, the fusion protein could be recognized by PAb -L5 raised against a Chinese isolate CTV -LS.
出处
《青岛农业大学学报（自然科学版）》
2009年第2期139-142,共4页
Journal of Qingdao Agricultural University(Natural Science)
关键词
柑橘衰退病毒
外壳蛋白
基因克隆
原核表达
citrus tristeza virus
coat protein
gene clone
prokaryotic expression
