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| 传染性喉气管炎病毒河南株ILTV-CG GX基因的克隆与序列分析 | |
| 引用本文： | 陈红英,崔保安,李新生,杨明凡,赵丽,方剑玉.传染性喉气管炎病毒河南株ILTV-CG GX基因的克隆与序列分析[J].华北农学报,2007,22(4):29-32. |
| 作者姓名： | 陈红英 崔保安 李新生 杨明凡 赵丽 方剑玉 |
| 作者单位： | 1. 河南农业大学,牧医工程学院,河南,郑州,450002;河南省动物性食品安全重点实验室,河南,郑州,450002 2. 河南农业大学,牧医工程学院,河南,郑州,450002 3. 南京农业大学,兽医学院,江苏,南京,210095 |
| 基金项目： | 国家“十五”食品安全重大攻关专项(2001BA804A30-11) |
| 摘 要： | 根据GenBank中收录的鸡传染性喉气管炎病毒(ILTV)GX基因核苷酸序列,设计并合成了1对引物,应用PCR技术扩增ILTV河南株(ILTV-CG)GX基因。将ILTV-CG接种10日龄鸡胚,提取ILTV基因组DNA进行PCR扩增。将回收的PCR产物与pGEM-T Easy载体进行连接,转化JM109感受态细胞,经蓝白斑筛选,对菌液PCR和酶切鉴定为阳性的菌株进行测序。测序结果表明,所克隆的GX基因全长为950 bp,含有一个开放阅读框,编码299个氨基酸的多肽。推导的氨基酸有4个潜在的N-糖基化位点,有11个与二硫键形成有关的半胱氨酸。序列分析表明,克隆的ILTV-CGGX基因与GenBank中发表的ILTV-SA2株GX基因核苷酸同源性为97.2%,变异点为第124位插入1个碱基G,第136位又插入2个碱基C,从而引起该毒株GX基因推导的氨基酸序列在第33位插入1个亮氨酸,还存在多处氨基酸发生变化,氨基酸同源性为95.3%。GX基因的成功克隆为构建ILTVGX基因缺失的病毒活载体奠定基础。 |
| 关 键 词： | 传染性喉气管炎病毒 GX基因 聚合酶链反应 序列分析 |
| 文章编号： | 1000-7091（2007）04-0029-04 |
| 修稿时间： | 2006-11-04 |
Cloning and Sequence Analysis of GX Gene of Chicken Infectious Laryngotracheitis Virus Henan Isolate ILTV-CG |
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| CHEN Hong-ying,CUI Bao-an,LI Xin-sheng,YANG Ming-fan,ZHAO Li,FANG Jian-yu.Cloning and Sequence Analysis of GX Gene of Chicken Infectious Laryngotracheitis Virus Henan Isolate ILTV-CG[J].Acta Agriculturae Boreali-Sinica,2007,22(4):29-32. | |
| Authors: | CHEN Hong-ying CUI Bao-an LI Xin-sheng YANG Ming-fan ZHAO Li FANG Jian-yu |
| Institution: | 1. College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China ; 2. Animal Food Safety Key Laboratory of Henan Province, Zhengzhou 450002, China; 3. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China |
| Abstract: | One pair of primers was designed according to the nucleotide sequence of chicken infectious laryngotracheitis virus(ILTV) GX gene from Genbank.The ILTV-CG strain was inoculated into 10-day embryonated eggs,which were used to extract the genome DNA for PCR amplification.The PCR products were purified,and then cloned into pGEM-T Easy vector.By screening of blue-white spot,the plasmid PCR and enzyme digestion,positive strains were sequenced.The results indicated that the amplified GX gene fragment was 950 bp,including an open-reading frame and encoding 299 amino acid residues that contain four potential N-glycosalation sites and eleven cysteines.The sequence analysis results showed a 97.2% similarity between GX gene ILTV-CG and ILTV-SA2 and 95.3% between their putative amino acid sequences.This study paved the way for preparation of recombinant vaccine of ILTV. |
| Keywords: | Infectious laryngotracheitis virus GX gene Polymerase chain reaction Sequence analysis |
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