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肛防御素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.
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 GenBank^TM(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.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2008年第3期61-65,共5页
Journal of South China Agricultural University
基金
广东省科技计划项目(2004A20106001
2007A020400006)