| 伪狂犬病病毒蛋白激酶基因部分序列的克隆和序列测定及转移质粒载体的构建 |
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| 引用本文： | 黄伟坚,陈德胜,姜焱,芦银华,张雪莲,陈溥言.伪狂犬病病毒蛋白激酶基因部分序列的克隆和序列测定及转移质粒载体的构建[J].中国预防兽医学报,2003,25(4):241-244. |
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| 作者姓名： | 黄伟坚 陈德胜 姜焱 芦银华 张雪莲 陈溥言 |
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| 作者单位： | 1. 南京农业大学,农业部动物疫病诊断和免疫重点开放实验室,江苏,南京,210095;广西大学,动物科学技术学院,广西,南宁,530005
2. 南京农业大学,农业部动物疫病诊断和免疫重点开放实验室,江苏,南京,210095 |
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| 摘 要： | 以伪狂犬病病毒SH株细胞培养物为模板，用PCR方法扩增蛋白激酶(PK)基因部分片段，克隆到pCDNA3中，序列测定显示所扩增的PK基因片段长907bp，与PRVNIA-3株的核苷酸同源性为99．6％。把PK基因克隆到pUCl9上，利用PK基因中间的SalI酶切位点，插入带有绿色荧光蛋白(EGFP)的基因表达盒，构成了含EGFP的转移载体，为今后研究PK基因的功能和构建PK缺失株奠定了基础。
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| 关 键 词： | 伪狂犬病病毒 蛋白激酶 PCR扩增 绿色荧光蛋白 转移载体 |
| 文章编号： | 1008-0589(2003)04-0241-04 |
| 修稿时间： | 2002年9月16日 |
Cloning and sequence analysis a part of protein kinase of pseudorabies virus and construction of PK-deleted-transfer vector |
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| HUANG Wei_jian ,CHEN De_sheng ,JIANG Yan ,LU Yin_hua ,ZHANG Xue_lian ,CHEN Pu_yan.Cloning and sequence analysis a part of protein kinase of pseudorabies virus and construction of PK-deleted-transfer vector[J].Chinese Journal of Preventive Veterinary Medicine,2003,25(4):241-244. |
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| Authors: | HUANG Wei_jian CHEN De_sheng JIANG Yan LU Yin_hua ZHANG Xue_lian CHEN Pu_yan |
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| Institution: | HUANG Wei_jian 1,2,CHEN De_sheng 1,JIANG Yan 1,LU Yin_hua 1,ZHANG Xue_lian 1,CHEN Pu_yan 1 |
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| Abstract: | The part of protein kinase gene in Us region of pseudorabies virus SH strain was amplified by polymerase chain reaction(PCR) and cloned in pCDNA3, its nucleotide sequence is 907 bp. The homology of PK gene nucleotide sequence of PRV SH strain with PRV NIA_3 strain is 99.6 %. The PK gene was cloned in pUC19 and cut with restriction enzyme SalI ,then the EGFP expression cassette was inserted into this site and the PK_ transfer vector containing EGFP expression cassette has been constructed. This result laid foundation to study the function of PRV PK gene and the development of engineered PK_deleted PRV strain. |
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| Keywords: | pseudorabies virus protein kinase PCR amplification EGFP transfervector |
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