广西黄鸡β-防御素基因的克隆、序列分析与组织分布

Cloning,Sequencing and Tissue Distribution of β-Defensin Gallinacin Genes in Guangxi Yellow Chicken

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.