摘要
目的了解禽流感病毒（AIV）在感染小鼠前后表面蛋白编码基因的特点及变异情况。方法在A／Goose／Guangdong／NH／2003（H5N1）感染小鼠肺组织的第12小时和第9天各分离1株AIV病毒，通过鸡胚增殖后提取RNA，反转录合成cDNA，经PCR扩增和产物纯化构建重组质粒，用双脱氧链终止法进行核苷酸序列测定并进行基因特性分析。结果3株病毒HA基因的核苷酸序列同源性为99．6％～99．8％，推导氨基酸序列的同源性为99．3％～99．6％。3株病毒NA基因的核苷酸序列同源性为99．8％-99．9％，均为同义突变。M基因未发生任何变异。结论AIV感染小鼠后HA基因出现变异，NA和肘基因未出现有意义的突变。
Objective To investigate the variations of surface protein genes of avian influenza virus （AIV） before and after infecting mouse. Methods Mouse lung tissue was infected with A/Goose/ Guangdong/NH/2003（ H5N1 ） and the virus was isolated 12 hours and 9 days after replication in lung tissue of mouse. The isolated strains were amplified in embryonated chicken eggs, and the virion RNA was transcribed into cDNA by reverse transcriptase. After amplification and purification, dideoxy-mediated chain termination was performed to detect synthetic oligonucleotide primers and DNA sequence was analyzed. Results The homology of nucleotide sequenee for HA gene of three isolated strains was 99.6% - 99.8% , and that of amino acid sequences was 99.3% - 99.6%. The homology of nucleotide sequence for NA gene of three strains was 99.8% - 99.9%, all of them were synonymous mutations. No variation was found in M gene. Conclusion After replication in mouse lung tissue, no significant mutation was found in the surface protein genes of AIV except some point mutations in HA genes.
出处
《中华临床感染病杂志》
CAS
2009年第2期93-97,共5页
Chinese Journal of Clinical Infectious Diseases
基金
广东省科技攻关项目（2003C31209）
广东省医学科研基金（A2006212）
广东省自然科学基金（06034508）
关键词
禽流感
基因分析
血凝素
神经氨酸酶
基质蛋白类
Avian influenza
Gene analysis
Hemagglutinins
Neuraminidase
Matrix protein
