摘要
在家蚕丝腺cDNA文库测序过程中,发现一个编码家蚕泛素结合酶的EST序列,利用3′RACE方法克隆了一个新的家蚕泛素结合酶基因cDNA全长序列,命名为BmUCE2 I(GenBank登录号为DQ219874)。家蚕BmUCE2 I基因全长cDNA由465 bp的开放阅读框序列(ORF)、97 bp的5′端非翻译区序列(5-′UTR)和237 bp的3′端非编码区序列(3-′UTR)组成,其编码的154个氨基酸与其他真核生物间具有较高的同源性。利用BmUCE2 I的EST片断作探针,通过筛选家蚕噬菌体基因组文库,获得了家蚕BmUCE2 I基因组序列和5′调控序列。BmUCE2 I基因由4个外显子和3个内含子组成,在5′端上游调控区域没有类似TATA盒元件,但在-219^-268 bp的区域存在一个50bp的启动子序列,此外还存在CF2-Ⅱ、FTZ、DFD、BRCZ2、DL、STAT、PRD-HD等多个转录因子结合位点。家蚕泛素结合酶新基因的克隆、基因结构及5′调控区的分析为进一步研究泛素蛋白水解酶复合通路相关基因的调控规律提供了重要依据。
According to the sequence of silkeworm EST generated by the large-scale cDNA sequencing, which is highly homologous to the ubiquitin conjugating enzyme （E2）, a full-length cDNA was cloned from the silkworm by 3′RACE, named as BmUCE21 （GenBank accession number DQ219874）. It contains an ORF of 465 bp , 97 bp nucleotide sequence in 5′UTR （untranslated region） and 237 bp nucleotide sequence in 3′ UTR （untranslated region）. It encodes 154 amino acids and shares highly homologenity to that of eukaryotic organism previously reported. By screening the λ phage genomic library derived from the silkworm, the BmUCE21 gene was cloned and sequenced. The BmUCE21 gene contains 4 exons and 3 introns and the putative promoter was between -219 bp - -268 bp of 5＇upstream sequence relative to translation start site. The BmUCE21 promoter contains no TATA box, but potential binding sites for the transcription factors were detected in both forward and reverse orientation ： CF2-Ⅱ ,FTZ,DFD,BRCZ2 ,DL,STAT,PRD-HD. The results of cDNA cloning, genomic structure and 5＇upstream analysis of the novel ubiquitin conjugating enzyme gene from silkworm provided a basis for relative gene control of Ubiquitin-Proteasomes Pathway.
出处
《蚕业科学》
CAS
CSCD
北大核心
2005年第4期439-443,共5页
ACTA SERICOLOGICA SINICA
基金
国家自然科学基金项目(编号30370805B30080023)
国家重点基础研究发展计划"973"项目(编号2005CB121003)
浙江省自然科学基金项目(编号Y304352)
关键词
家蚕
泛素结合酶基因
基因克隆
启动子
Bombyx mori
Ubiquitin conjugating enzyme gene
Gene cloning
Promoter
