| 利用交换接头技术构建大麦条纹花叶病毒新疆株RNA_2的全长cDNA克隆 |
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| 引用本文： | 刘中大,彭学贤,曲士绵,张鹤龄,莽克强,Wing Lam Sung.利用交换接头技术构建大麦条纹花叶病毒新疆株RNA_2的全长cDNA克隆[J].病毒学报,1991(4). |
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| 作者姓名： | 刘中大 彭学贤 曲士绵 张鹤龄 莽克强 Wing Lam Sung |
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| 作者单位： | 内蒙古大学生物系,中国科学院微生物研究所,中国科学院微生物研究所,内蒙古大学生物系,中国科学院微生物研究所,Institute for Biological Science,National Research Council of Canada,Ortawa |
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| 摘 要： | 本文采用双股交换接头(Crossover linker)技术对已建立的大麦条纹花叶病毒新疆株(BSMV-XJ)RNA_2 cDNA重组质粒pUBS_(112)进行修饰。使cDNA两端不必要的poly(dG):poly(dC)尾准确地缺失,同时补上了cDNA中相对于RNA_2 5’端区缺少的三个核苷酸TAA,并在cDNA末端插入了预定的限制酶切顺序。通过原位杂交筛选、酶切图谱分析、cDNA两端序列测定等手段,证明已获得BSMV-XJ RNA_2组分的全长cDNA克隆。
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| 关 键 词： | 大麦条纹花叶病毒 全长cDNA克隆 交换接头 |
CONSTRUCTION OF FULL-LENGTH RNA_2 cDNA CLONE OF BARLEY STRIPE MOSAIC VIRUS SINGJIANG STRAIN ( BSMV-XJ ) BY THE CROSSOVER LINKER TECHNIQUE |
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| Liu Zhongda Peng Xuexian Qu Shimian Zhang Heling Mang Keqiang Wing Lam Sung Institute of Microbiology,Academic Sinica,Beijing.CONSTRUCTION OF FULL-LENGTH RNA_2 cDNA CLONE OF BARLEY STRIPE MOSAIC VIRUS SINGJIANG STRAIN ( BSMV-XJ ) BY THE CROSSOVER LINKER TECHNIQUE[J].Chinese Journal of Virology,1991(4). |
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| Authors: | Liu Zhongda Peng Xuexian Qu Shimian Zhang Heling Mang Keqiang Wing Lam Sung Institute of Microbiology Academic Sinica Beijing |
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| Institution: | Liu Zhongda Peng Xuexian Qu Shimian Zhang Heling Mang Keqiang Wing Lam Sung Institute of Microbiology,Academic Sinica,BeijingDepartment of Biology,The Inner Mongolia University,HohhotInstitute for Biological Sciences,National Research Council of Canada,Ottawa |
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| Abstract: | In order to obtain full-length cDNA,the "duplex crossover linker technique" was used to modify the 3'-and 5'-end of the cDNA insert of BSMV-XJ RNA2 in recombinant plasmid, pUBS112, which had been previously established ( by making deletions and insertions ) . The unwanted poly ( dG ) . poly ( dC ) tails of the both sides of the cDNA segment were precisely deleted.The predetermined restriction sequences and the lacked three nucleotides TAA in original pUBS112 which correspond to 5'-terminal of BSMV-XJ RNA2 were added at the deleted sites. The mutant was selected by in situ hybridization with r-32p-labelled synthetic crossover linkers as probe. The restriction map and sequence of the selected clone, pUfBS4, were compared with those of pUBSll2 and RNA2 of BSMV type strain. The results showed that recombinant fragment in pUfBS was full-length RNA2 cDNA of BSMV-XJ. |
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| Keywords: | Barley stripe mosaic virus Full-length cDNA cloneCrossover linker |
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