摘要
本文论述了蓖麻蚕卵黄原蛋白cDNA迅速克隆化的方法。SDS-PAGE鉴定的蓖麻蚕卵黄原蛋白由大小二个亚基构成,求出的分子量大亚基为180kDa,小亚基为45kDa,从解析的蓖麻蚕卵黄原蛋白氨基酸序列推算的分子量大亚基为161.06kDa,小亚基为40.53kDa,如果考虑到翻译后的修饰,这与SDS-PAGE求出的分子量是吻合的。蓖麻蚕卵黄原蛋白cDNA是由5788pb构成,一个ORF为5337个碱基,编码了1779个氨基酸。在信号肽的15个氨基酸残基中,有12个是疏水性氨基酸残基。这与其他昆虫卵黄原蛋白信号肽区域的疏水性分析是一致的。蓖麻蚕卵黄原蛋白N-linkedglycosylationsite的分布与家蚕、柞蚕和天蚕不同,3处N-linkedglycosylationsite存在于大亚基里,2处存在于多聚丝氨酸区域上流的小亚基里。另外,我们发现在所解析的柞蚕、天蚕和蓖麻蚕卵黄原蛋白氨基酸序列的C-末端区域里DGQR、GICG功能部位及其后的6个半胱氨酸都完好地保存。
This paper states a simpl e and quick method of cloning and analysis of amino acid sequence of vitellogeni ns in eri-silkworm( Philosamia cynthia ricini ). Through SDS-PAGE electroph oresis vitellogenins in Philosamia cynthia ricini is confirmed to be made of two subunits of 180kDa and the 45kDa respestively. According to analysis of am ino acid sequence, the big subunit is 161.06kDa and the small one is 40.53kDa,i f modification of post-translation is under consideration, which is identical t o that of SDS-PAGE electrophoresis. The cDNA of vitellogenins in Philosamia cynthia ricini consists of 5788bp, one ORF is 5337 bases which codes 1779 am ino acids. There are 12 hydrophobic residues among fifteen amino acid residues i n the s ignal peptide, which is identical to that of Vg in other insects. The distributi on of N-linked glycosylation site of Vg in Philosamia cynthia ricini is d if ferent from Bombyx mori, Antheraea pernyi, Antheraea yamamai, which exists i n big subunit in three sites and in polyserine domain upstream small subunit in two sites. Besides, in the C-ends domains of amino acids of Vg in Antheraea p ernyi, Antheraea yamamai, Philosamia cynthia ricini , DGQR and GICG structur al unit and six subsequent cysteines are well preserved. [WT5HZ]
出处
《激光生物学报》
CAS
CSCD
2003年第1期12-17,共6页
Acta Laser Biology Sinica
基金
安徽省自然科学基金资助项目(01041201)
安徽省教育厅基金资助项目(2001kj085)
关键词
蓖麻蚕
卵黄原蛋白
CDNA
eri-silkworm( Philosamia cynthia r icini.)
vitellogenin(Vg)
cDNA
