【摘要】 用设计的特异性引物,采用RT-PCR方法扩增到广西黄鸡β-防御素基因gallinacin-1(简称Gal-1)~gallinacin-13(简称Gal-13).通过克隆、测序获得这13个基因的cDNA核苷酸序列,并提交到GenBank(Gal-1~Gal-13基因的注册号为:DQ858297~DQ858310).比较分析广西黄鸡13种β-防御素Gal-1~Gal-13基因分别与GenBank中注册的防御素基因氨基酸序列的相似性,其氨基酸的相似性均在95.5%~100%之间.利用DNAStar软件对所获得的13种广西黄鸡β-防御素基因进行系统发育树分析,结果Gal-1与Gal-5、Gal-12、Gal-2在同一分支上,Gal-6、Gal-7、Gal-9、Gal-8在同一分支上,Gal-10、Gal-11、Gal-4在同一分支,Gal-13独自为一分支.同样用RT-PCR方法分析了广西黄鸡β-防御素Gal-1~Gal-13基因在不同组织中的分布,结果发现:Gal-1、Gal-2、Gal-4、Gal-5、Gal-6、Gal-7、Gal-10的表达分布非常广泛,Gal-3、Gal-8、Gal-9、Gal-11、Gal-12、Gal-13的分布较少.更多还原

【Abstract】 With specific primers,thirteen chicken β-defensin gallinacin-1(Gal-1)～gallinacin-13(Gal-13) cDNA fragments,were amplified by reverse transcription-polymerase chain reaction(RT-PCR) from Guangxi Yellow chicken.All of the cDNA fragments covered the full-length of the open reading frame,with the exception of Gal-3.After being sequenced,13 cDNA fragments were identified and deposited to GenBankTM(accession numbers DQ858297~ DQ858310).The results of sequence comparison with the β-defensin gene sequences published in GenBankTM indicated that amino acid similarity of the 13 chicken β-defensin genes ranged from 95.5% to 100%.Analysis by DNAstar showed that Gal-1,Gal-5,Gal-12 and Gal-2 lay at the same embranchment;Gal-6,Gal-7,Gal-9 and Gal-8 lay at the same one;Gal-10,Gal-11 and Gal-4 lay at the same one;Gal-13 lay at the embranchment independently.Different mRNA distribution of genes in different tissues,analyzed by RT-PCR,indicated that Gal-1,Gal-2,Gal-4,Gal-5,Gal-6,Gal-7 and Gal-10 were widely expressed in tissues,while Gal-3,Gal-8,Gal-11,Gal-12 and Gal-13 had limited tissue distribution.更多还原