抗禽流感病毒免疫相关基因的筛选与鉴定

 【目的】研究抗禽流感病毒免疫的分子机制，发现新的抗禽流感病毒免疫相关基因，为抗禽流感转基因育种的研究奠定基础。【方法】随机选取40只30日龄AA白羽肉鸡，其中20只鸡注射禽流感H5亚型禽流感灭活疫苗，另20只未注射疫苗鸡作为对照，在注射后第9天取脾脏保存在液氮中。应用mRNA差异显示技术，结合反向northern dot blot鉴定技术，来筛选注射疫苗组和未注射疫苗组的差异表达基因。【结果】筛选出13条差异表达EST，其中未注射疫苗组4条，注射组9条表达量增高。应用BLAST工具将EST对核酸数据库nr和dbEST中所有序列进行了同源性分析。DD3、DD4、DD6、DD9、DD11、DD12与已有核酸数据库中的基因克隆或EST具有较高相似性，为已知的EST，但功能未知。DD10、DD13与鸡核糖体蛋白L7a基因具有很高的相似性。DD1、DD2、DD5、DD8为新的EST，提交到GenBank，分别获得登录号为EB714185、EB714186、EB714187、EB714188。【结论】本研究筛选出的鸡核糖体蛋白L7a基因和其它EST对应的未知功能基因可以作为抗禽流感病毒研究中的候选基因，其具体的功能有待今后进一步研究。

Screening and Identification of Anti-Avian Influenza Virus Immune-Related Genes

【Objective】This study was carried out to probe the molecular mechanism of inactivated influenza virus vaccines immunity and find new anti-avian influenza virus genes. In addition, this study may provide candidate genes for producing anti-avian influenza virus transgenic chickens. 【Method】 40 Abar acer chickens(AA) were selected randomly in this study, 20 of which were injected H5 inactivated influenza virus vaccines ,the other 20 were controls. The 40 chickens were killed. Spleen was saved in liquid nitrogen. mRNA differential display PCR and reverse Northern dot bloting were used to find differential expressed genes between injected group and uninjected group. 【Result】13 ESTs were found via DDRT-PCR、reverse northern dot blot, 9 of which in injected group, the other 4 were in control. All 13 ESTs were compared with nucleotide sequences deposited in the nr database and the dbEST database of Genbank via BLASTn tool. DD10 and DD13 were found similar to gallus mRNA for ribosomal protein L7a. DD3, DD4, DD6, DD9, DD11 and DD12 ESTs had their highly similar nucleotide sequences in nucleotide databases but with unknown functions. The other 4 ESTs had no significant similarity with existing genes or ESTs and were regarded as new ESTs. The 4 new ESTs were submitted to GenBank(Accession number: EB714185, EB714186, EB714187, EB714188). 【Conclusion】 Gallus ribosomal protein L7a is probably connected in Avian Influenza vaccines immunologic process and anti-virus process but its exact mechanism needs further study.