狮头鹅α-干扰素基因的克隆与遗传分析  

Clonging and Sequence Analysis of the Interferon-α Gene of Shitou Goose

作　　者：郑晓灵[1] 卢曦[1] 刘艳芬[1] 陈绍红[2] 杨乐乐[1] 刘铀[2] 

机构地区：[1]广东海洋大学农学院,湛江524088 [2]广东海洋大学生化中心,湛江524088

出　　处：《中国畜牧兽医》2008年第6期29-33,共5页China Animal Husbandry & Veterinary Medicine

摘　　要：从狮头鹅外周血细胞中提取基因组DNA,采用巢式PCR方法扩增α-干扰素基因片段（d-Go-IFN-α）;同时从新城疫病毒（NDV）刺激培养外周血淋巴细胞提取总RNA,用RT-PCR方法扩增α-干扰素基因片段（m-Go-IFN-α）,将上述扩增片段分别克隆到pGEM-T载体并进行序列测定。结果表明,α-干扰素基因全长均为576 bp,编码192个氨基酸,比较d-Go-IFN-α和m-Go-IFN-α的核苷酸序列,可见155位核苷酸由A突变为G,相应地52位氨基酸由M突变为R,提示狮头鹅α-干扰素基因存在单链核苷酸多态性;狮头鹅与其它家禽品种α-干扰素基因的核苷酸序列同源性介于41.2%~98.3%之间,其推导的氨基酸序列同源性为13.0%~96.4%,不同鹅品种的α-干扰素具有极为相似的二级结构。With nested transcription polymerrase chain reaction,the interferon-α gene fragment of shitou goose（d-Go-IFN-α） was amplified from genome DNA extracted from peripheral blood cells（PBCs）.At the same time,the total RNA was extracted from peripheral blood lymphocyte（PBLs） stimulated with Newcasttle disease virus（NDV） in vivo,and transcription polymerrase chain reaction（RT-PCR） was used to amplify the interferon-α gene fragment of shitou goose（m-Go-IFN-α）,then both of the amplified fragments were cloned into pGEM-T easy vector and sequenced.The results showed that both of the d-Go-IFN-α and m-Go-IFN-α were both consisted of 576 necleotides,coding for 192 amino acids.Compared with the nucleotide sequences of d-Go-IFN-α and m-Go-IFN-α,a variation A→G at the 155th site was observed,and the 52th amino acid was mutated from M to R accordingly,which implied that there was single nucleotde polymorphism in IFN-α sequence.of Shitou goose.The homolony of nucleotide and the deduced amino acid sequence among different poultry breeds were 41.2 % to 98.3% and 13.0% to 96.4% respectively.The secondary structure of the IFN-α were almost the same among different goose breeds.

关 键 词：狮头鹅 Α-干扰素 克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]