摘要
目的 克隆家鸽、鹌鹑与人类发育缺陷增强因子 (enhancerofrudimentary ,e(r) )基因。方法 依据来源于家鸽的e(r)基因同源EST片段设计引物。提取家鸽与鹌鹑脑组织RNA ,逆转录为cDNA。通过RACE方法克隆家鸽e(r)基因全长序列。依据家鸽序列设计引物 ,通过PCR方法扩增鹌鹑e(r)同源基因的编码区。依据人类e(r)基因序列设计引物 ,利用人类脑组织RNA扩增人类e(r)基因编码区。测定 3种基因的DNA序列 ,并进行同源性比较。结果 自家鸽克隆得到 776bp的e(r)基因全长cDNA序列、自鹌鹑与人类克隆得到 312bp的e(r)基因编码区序列。经同源性比较证实 ,三者序列高度同源 ,其氨基酸序列同源性分别为 99% ,99%和 10 0 %。结论 根据e(r)基因的引物已成功克隆出家鸽、鹌鹑和人类的e(r)基因 ,且同源性很高。
Objective To clone the enhancer of rudimentary gene from pigeon,partridge and human.Methods Primers were designed by EST segment rooted in enhancer of rudimentary(e(r)) gene of pigeon.Total RNA of pigeon and partridge were purified from brain and reverse transcripted to cDNA.Full length cDNA of e(r) gene of pigeon was cloned by rapid amplification of cDNA ends(RACE).Primers were designed by e(r) gene sequence of pigeon and used to amplify the coding region of e(r) homologous gene of partridge.Primers were designed by human e(r) gene sequence and amplified the coding region of human e(r) gene using human brain RNA.Three genes were cloned into vector, sequenced and their homology were compared.Results 776 bp full length e(r) gene cDNA of pigeon and 315 bp e(r) gene coding region of partridge and human were obtained.It has been confirmed that high degree of conservation existed among e(r) gene of pigeon, partridge and human.The amino acid sequences of pigeon,partridge and human proteins are 99%,99% and 100% identical respectively.Conclusion Cloning e(r) gene of pigeon, partridge and human by primer of e(r) gene was successfully done with high conservation.
出处
《同济大学学报（医学版）》
CAS
2004年第5期378-380,384,共4页
Journal of Tongji University(Medical Science)
