一步法克隆传染性法氏囊病病毒前体多聚蛋白基因  

Cloning of Precursor Polyprotein Gene of Infectious Bursal Disease Virus by One Step

作　　者：刘存仁[1] 梁志清[2] 

机构地区：[1]山东大学生命科学学院,济南250100 [2]香港大学动物学系

出　　处：《病毒学报》2001年第2期180-182,共3页Chinese Journal of Virology

基　　金：国家教育部留学回国人员基金

摘　　要：Very virulent infectious bursal disease virus(vvIBDV)was isolated from chicken bursa and then the virus double stranded RNA was extracted After denaturation of dsRNA by heat in the presence of primers,the cDNA was synthesized by use of a reverse transcriptase lacking RNase H activity The RNA component of RNA cDNA hybrids was digested by RNase H By using an optimized PCR,a 3 05kb DNA fragment coding for precursor polyprotein of IBDV was produced in one step,which was inserted into pcDNA3 1(+) vector Two recombinant plasmids (pPP1 and pPP2)were screened and identified from eight XL1 blue colonies Partial sequencing for pPP1 plasmid indicated that the precursor polyprotein gene for IBDV was cloned successfully The method can simplify greatly the procedure to clone precursor polyprotein gene of IBDVVery virulent infectious bursal disease virus(vvIBDV)was isolated from chicken bursa and then the virus double stranded RNA was extracted After denaturation of dsRNA by heat in the presence of primers,the cDNA was synthesized by use of a reverse transcriptase lacking RNase H activity The RNA component of RNA cDNA hybrids was digested by RNase H By using an optimized PCR,a 3 05kb DNA fragment coding for precursor polyprotein of IBDV was produced in one step,which was inserted into pcDNA3 1(+) vector Two recombinant plasmids (pPP1 and pPP2)were screened and identified from eight XL1 blue colonies Partial sequencing for pPP1 plasmid indicated that the precursor polyprotein gene for IBDV was cloned successfully The method can simplify greatly the procedure to clone precursor polyprotein gene of IBDV isolates

关 键 词：鸡 传染性法氏囊病病毒 前体多聚蛋白 基因 克隆 

分 类 号：S858.315.3[农业科学—临床兽医学] S852.659.4[农业科学—兽医学]