鸭IGF-IR基因部分序列克隆、鉴定及在胚胎期胸肌组织中的差异表达分析

Cloning,Sequence Identification and Expression Analysis of IGF-IR in Breast Muscle at Different Embryonic Period in Ducks

为了克隆分析鸭IGF-IR基因部分的cDNA序列,并研究该基因的组织表达规律。通过同源性比对,克隆了IGF-IR基因cDNA部分片段,并运用生物信息学方法分析了不同物种序列进化关系,运用荧光绝对定量PCR技术检测了IGF-IR mRNA在肉用型品种高邮鸭和蛋用型品种金定鸭胚胎期不同发育时期胸肌中的表达。序列分析结果表明:克隆得到的鸭IGF-IR基因cDNA270bp片段长度,其核苷酸序列与鸡、人、小鼠、大鼠、猪、牛的IGF-IR基因相应核苷酸序列的同源性分别为95%、79%、78%、77%、78%、76%;荧光绝对定量结果表明:IGF-IR基因在胸肌中呈"波浪形"表达,在21胚龄表达量最高,与17、25、27胚龄和7日龄差异显著(P<0.05);IGF-IR基因在品种间表达量除21胚龄时差异显著外(P<0.05),其余各胚龄差异均不显著(P>0.05),表明IGF-IRmRNA的表达在所研究品种胚胎期的胸肌中相对稳定。

The present experiment was conducted to obtain and analyze partial cDNA sequence of duck Insulin-like growth factor I receptor gene （IGF-IR）,and to study the tissue expression characterization of IGF-IR mRNA in ducks.Partial sequence of IGF-IR cDNA was cloned by homologous alignment analysis,the sequence was analyzed by bioinformatics method,and its absolute mRNA expression was detected by quantitative RT-PCR in breast muscle at different embryonic period meat-type duck breed-Gaoyou ducks and egg-type duck breed-Jinding ducks.Sequence analysis showed that the 270 bp of CDS region of duck IGF-IR gene shared 95%,79%,78%,77%,78%,76% identity with those of Gallus gallus,Homo sapiens,Mus-musculus,Rattus novegicus,Sus domesticus and Bos Taurus,respectively.The results of quantitative RT-PCR indicated that the expression of duck IGF-IR showed ＂ wave ＂ trend at different embryonic period,the highest level appeared at 21 embryo age,which was significant higher than that of 17,25,27 embryo age and 7was significant at 21 embryo age （P〈0.05）,and the differences were not significant at other embryo age （P〈0.05）.