家蝇卵黄蛋白基因的克隆与结构序列分析

Cloning and Sequence Analysis of Musca domestica Yolk Protein Gene

家蝇卵黄蛋白基因编码的卵黄蛋白是家蝇胚胎发育的重要营养来源 .根据 3种家蝇卵黄蛋白cDNA保守序列设计引物 ,用PCR技术从家蝇基因组DNA中扩增到大小为 76 8bp的mdYP1基因的部分DNA片段 .经地高辛标记成特异性探针 ,从构建的家蝇基因组文库中筛选出一个阳性克隆 ,并从该克隆中分离到大小为 3991bp的mdYP1基因组基因 .序列分析显示 ,该基因组序列含有约1 6kb的 5′ 上游区和 1 0kb的 3′ 下游区 ,编码区由一个 6 1bp的内含子和大小分别为 2 2 2bp和10 2 8bp的 2个外显子组成 .5′ 上游区含有典型的CAAT TATA盒 .

Musca domestica yolk protein (mdyp) genes encode the major nutritional polypeptides deposited in developing oocytes for subsequent utilization during embryogenesis. About a 4 0 kb genomic gene encoding Musca domestica yolk protein 1(mdyp1) was cloned by screening Musca domestica genomic library constructed in EMBL3 vector with a mdyp1 DNA fragment obtained by PCR as probe labeling with Digoxigen. Sequence analysis revealed that it contained about 1 6 kb 5′ upstream region and about1 0 kb 3′ downstream region. Encoding region was composed of a 61 bp intron, a 222 bp exon 1 and a 1028 bp exon 2. 5′ upstream region possesses typical CAAT/TATA boxes.