猪RPS3基因的克隆和表达分析  

Cloning and Expression Analysis of Porcine RPS3 Gene

作　　者：杨秀芹[1] 郭丽娟[1] 关庆芝[1] 刘惠[1] 刘娣 

机构地区：[1]东北农业大学动物科技学院,哈尔滨150030

出　　处：《畜牧兽医学报》2007年第7期630-635,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：黑龙江省杰出青年基金(JC-05-19);黑龙江省科技攻关项目(GB05B106)

摘　　要：RPS3在体内具有多种功能,为了进一步揭示其结构与功能的关系,探索其编码区作为系统发育标记的可行性,本研究应用生物信息学、RT-PCR和3-′RACE方法首次克隆了猪RPS3基因(GenBank登录号:DQ660373)并进行了序列分析,同时利用半定量RT-PCR方法分析了RPS3基因的时空表达规律。序列分析表明RPS3开放读码框全长为732 bp,编码243个氨基酸残基的多肽链,含有核糖体蛋白S3标签,构建的系统发育树显示聚类结果与动物学上的分类一致。半定量RT-PCR分析表明RPS3在所检测的10种组织中均有较高水平的表达,心脏中的表达量最高,肺脏中的表达量最低,其它组织间差异不大;在仔猪中表达量高,随着日龄的增加表达量不断下降,在3月龄以后的猪中表达量趋于平稳。以上结果说明RPS3基因CDS适合构建种间的系统发育树,mRNA的表达水平与细胞的生长代谢速度有关。In order to further elucidate the structure-function relationship of RPS3 which has multifunction in vivo and to reveal the possibility of its CDS as a phylogenetic marker, the RPS3 cDNA （GenBank accession No： DQ660373） was cloned using bioinformatics, RT-PCR and 3＇-RACE methods; and its expression profile was researched with semi-quantitative RT-PCR. Sequence analyses showed that the full ORF is 732 bp, encoding a polypeptide of 243 amino acids which ineludes the ribosomal protein S3 signature. The phylogenetic tree demonstrated similar topology with the zoology. Semi-quantitative RT-PCR analyses showed that the expression level is the highest in heart and the lowest in liver, and there are no significant difference among other tissues. With the age increasing, the expression level gradually decreases and becomes steady in pigs older than three months. The results made clear that the coding sequence of RPS3 fits to construct phylogenetic tree among species and its expression at mRNA level is related to growth and metabolism .

关 键 词：猪 RPS3基因 克隆 序列分析 半定量RT-PCR 

分 类 号：S828.2[农业科学—畜牧学]