| 猪细小病毒PPV-SD1株VP2全基因的克隆及原核表达 |
| |
| 引用本文： | 王金良,沈志强,谢金文,南松剑,管宇,郝静,庄金秋,刘吉山.猪细小病毒PPV-SD1株VP2全基因的克隆及原核表达[J].畜牧与兽医,2007,39(12):14-17. |
| |
| 作者姓名： | 王金良 沈志强 谢金文 南松剑 管宇 郝静 庄金秋 刘吉山 |
| |
| 作者单位： | 山东省滨州畜牧兽医研究院,山东省畜禽用蜂胶疫苗工程技术研究中心,山东,滨州,256600 |
| |
| 基金项目： | 山东省自然科学基金(Y2005D10) |
| |
| 摘 要： | 为研究猪细小病毒PPV-SD1分离株的VP2蛋白的结构与功能,自行设计引物,通过PCR扩增的方法,获得VP2全基因,克隆到pMD18-T载体中进行测序,然后亚克隆到pET30a(+)表达载体中,构建并筛选出阳性重组子,标记为pET30a-VP2。将阳性重组质粒转化进RosettaTM宿主菌中,通过改变IPTG浓度、诱导温度和诱导时间,使重组蛋白获得表达,并确定最佳诱导条件为:IPTG终浓度1.0 mmol/L、诱导温度37℃、诱导时间为3~4 h。经SDS-PAGE和Western-blotting分析表明该重组蛋白相对分子量约为68 ku,具有良好的免疫学活性。
|
| 关 键 词： | 猪细小病毒 VP2基因 基因克隆 原核表达 |
| 文章编号： | 0529-5130（2007）12-0014-04 |
| 收稿时间： | 2007-05-09 |
| 修稿时间： | 2007年5月9日 |
Cloning and prokaryotic expression of VP2 gene from porcine parvovirus PPV-SD1 strain |
| |
| WANG Jin-liang,SHEN Zhi-qiang,XIE Jin-wen,NAN Song-jian,GUAN Yu,HAO Jing,ZHUANG Jin-qiu,LIU Ji-shan.Cloning and prokaryotic expression of VP2 gene from porcine parvovirus PPV-SD1 strain[J].Animal Husbandry & Veterinary Medicine,2007,39(12):14-17. |
| |
| Authors: | WANG Jin-liang SHEN Zhi-qiang XIE Jin-wen NAN Song-jian GUAN Yu HAO Jing ZHUANG Jin-qiu LIU Ji-shan |
| |
| Abstract: | In order to study the structure and function of VP2 protein from porcine parvovirus PPV-SD1 strain,full-length VP2 gene was amplified by PCR,and then cloned into pMD18-T and sequenced.The recombinant plasmid pET30a-VP2 was constructed by subcloning into expression vector pET30a(+) and transformed into RosettaTM.VP2 gene was successfully expressed in host cells when induced at 37 ℃ with 1.0 mM IPTG for 3-4 h.This recombinant protein with a molecular weight of about 68 ku was approved to have immunological activity by SDS-PAGE and Western-blotting. |
| |
| Keywords: | porcine parvovirus VP2 gene cloning prokaryotic expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
