云南农业大学学报2013，Vol.28Issue(1)：32-35,4.DOI:10.3969/j.issn.1004-390X (n).2013.01.006
香石竹斑驳病毒外壳蛋白基因的原核表达及抗血清制备
Prokaryotic Expression and Antiserum Preparation of the Coat Protein Gene of Carnation mottle virus
摘要
Abstract
Carnation mottle virus (CarMV) is one of the important viruses infecting carnation. In this study, RNA was extracted from CarMV infected carnation, and coat protein (cp) gene was cloned into pET30a ( + ) and transformed into BL21 (DE3) pLysS. The optimal conditionsor the highest expres-sion of coat protein was expressing at 37℃ for 6 h and inducing with 1. 0 mmol/L IPTG. The expressed protein was purified with gradient SDS-PAGE and the antiserum against the protein was raised in Kun-ming mice. The result of Western-blot analysis confirmed that the antiserum reacted strongly and spe-cifically recombinant CarMV cp and CarMV infected samples. Forty-eight carnation samples in Kun-ming were tested with the antibody made in this experiment, of which 79. 2% of the samples were in-fected by CarMV. This study provides a good basis for massive CarMV cp antiserum preparation CarMV detection and interaction studies between CarMV and host plants.关键词
香石竹斑驳病毒/外壳蛋白基因/原核表达/抗血清/检测Key words
Carnation mottle virus/coat protein gene/prokaryotic expression/antiserum/detection分类
农业科技引用本文复制引用
李晓鹏,王连春,彭杰军,姬盼,李准,孔宝华,陈海如..香石竹斑驳病毒外壳蛋白基因的原核表达及抗血清制备[J].云南农业大学学报,2013,28(1):32-35,4.基金项目
云南省攻关项目(2006NG02). （2006NG02）
