香蕉条斑病毒CP区基因的克隆及其在大肠杆菌中的融合表达Cloning of Coat Protein Gene of Banana Streak Virus and its Fusion Expression in Escherichia coli
邱世明;庄军;刘志昕;
摘要(Abstract)：
依据锌指结构在BSVCP上的可能位置合成了2条引物,通过PCR扩增得到长约640 bp的目的片段;将目的片段与质粒pET28 b(+)连接,构建了含BSVCP区基因的融合蛋白原核表达载体pET28b-BSVCP;克隆测序以确定其阅读编码框的正确性,然后用正确的重组质粒转化大肠杆菌Rosetta(DE3),并诱导表达;经IPTG诱导后,出现一特异的约30 kDa融合蛋白质表达条带。
关键词(KeyWords)： 香蕉条斑病毒;外壳蛋白;原核表达
基金项目(Foundation): 国家自然科学基金资助项目(30660100);; 海南省教育厅高等学校科研资助项目(Hjkj200505)
作者(Author): 邱世明;庄军;刘志昕;
Email:
DOI:
参考文献(References)：
- [1]THOTTAPPILLY G,DAHAL G,LOCKHART B E L.Studies on a Nigerianis late of banana streakbadnavirus:I.Purification and enzyme linkeimmunosorbent assay[J].Ann Appl Biol,1998,132(2):253-261.
- [2]FRISON E A,SHARROCK S L.Banana streak virus:aunique virus Musa interaction[J].J AOOC A,1998(27):70.
- [3]LOCKHART B E.Purification and serology of a bacilliformvirus associated with banana streak disease[J].Phytopathology,1986,7(10):995-999.
- [4]DAHALET,DAHAL G,HUGHES J.Status of banana streak diseasein Africa:problems and future research needs[J].Integrated PestMngmt,1998,3:85-97.
- [5]DIEKMANN,PUTTER.Technical guidelinesfor the safe movement ofgermplasm[M].2nd edn.Rome:FAO/IPGRI.
- [6]HARPER G,HULL R.Cloning and sequence analysis of bananastreak virus DNA[J].Virus Genes,1978,17:271-278.
- [7]ETIENNE H Z,ORLENE G P,THOMAS H.The rice tungrobacilliform virus gene II product interacts with the coat proteindomain of the viral[J].Journal of Virology,2000,74(5):2073-2083.
- [8]古勤生.中国侵染葫芦科作物病毒的检测和小西葫芦黄化花叶病毒的变异性[D].北京:中国农业大学,2001.
- [9]SAM B J,FRITSCH E F,MANIATIS T.Molecular cloning:a laboratorymanual[M].New York:Cold Spring Harbor Laboratory Press,1989:885-889.