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当前位置: 首页 > 文章 > 版纳微型猪近交系INF-α基因克隆、序列分析及分子系统进化研究 西南农业学报 2012,25 (6)
Position: Home > Articles > Cloning, Sequence Analysis and Molecular Phylogeny Evolution of  TNF-α Gene in Banna Mini-pig Inbred Line (BMI) Southwest China Journal of Agricultural Sciences 2012,25 (6)

版纳微型猪近交系INF-α基因克隆、序列分析及分子系统进化研究

作  者：
赵跃;王锐;霍金龙;刘丽仙;霍海龙;朱国琪;曾养志
单  位：
云南农业职业技术学院畜牧兽医系;云南农业大学动物科学技术学院
关键词：
版纳微型猪近交系;TNF-α基因;序列分析;系统进化;组织表达
摘  要：
分别提取版纳微型猪近交系18种组织的RNA,反转录为cDNA,利用RT-PCR方法扩增肿瘤坏死因子(TNF-α)基因编码区全长序列,将纯化的片段与pMDl8-T载体连接,转化大肠杆菌DH5eL,筛选阳性克隆并测定其序列,利用生物信息学方法分析其序列特征,并与其他15个物种构建系统进化树。同时采用半定量RT-PCR方法分析TNF-α基因在版纳微型猪近交系18种组织中的表达情况。结果显示,版纳微型猪近交系TNF-α基因编码区全长699bp(GenBank登录号:JF831365),编码232个氨基酸。氨基酸序列同源性分析表明,与猫等12个物种具有80%以上的相似性,分子系统进化表明与牛、山羊、绵羊及海豚的关系较近。多组织半定量RT-PCR研究表明,TNF-αmRNA在版纳微型猪近交系的多个组织中均有表达,其中在中脑、卵巢、脾、肺、神经及胃中表达量较高。
译  名：
Cloning, Sequence Analysis and Molecular Phylogeny Evolution of  TNF-α Gene in Banna Mini-pig Inbred Line (BMI)
关键词：
Banna Mini-pig Inbred Line (BMI) % TNF-α gene% Sequence analysis% Phylogeny evolution% Tissue expression
摘  要：
The RNAs of 18 kinds of tissues from Banna Mini-pig Inbred Line (BMI) were extracted, separately and reversely transcribed into cDNA. The gene coding region sequence of tumor necrosis factor-alpha was amplified through RT-PCR, and then the purified product of PCR was ligated with a pMD18-T and transferred into the bacterium DH5α for replication. The positive clones were screened and sequenced. The sequence characters were analyzed by using bioinformatics method and phylogeny evolution tree was constructed with other fifteen species. Meanwhile, the semi-quantitative RT-PCR was used to analyze the expression profile of TNF-α gene in 18 kinds of tissues of BMI. The resuits showed that the coding region of TNF-α gene was 699 bp ( GenBank accession number:JF831365), and coded 232 amino acids. The amino acid homology analysis showed that it had over 80 % homology with 12 species, such as eat. Molecular phylogeny evolution indicated that it had a close relation with cattle, goat, sheep and dolphin. Semi-quantitative RT-PCR analysis of mRNA from different tissues indicted that TNF-α mRNA expressed in several tissues of BMI, and the expression level was higher in brain, ovary, spleen, lung, nerve fiber and stomach than other tissues.

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