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蜘蛛丝基因MaSp1在转基因家蚕中的表达分析
韩冷,张磊,中垣雅雄
0
(信州大学纤维学部应用生物系,上田 386-8567;信州大学纤维学部应用生物系,上田 386-8567; 安徽省农业科学院蚕桑研究所,合肥 230061)
摘要:
利用反向PCR、定量PCR以及免疫共沉淀等方法分析研究转基因家蚕的丝素蛋白及基因的结构和性质。结果表明:(1)在DNA 水平上,MaSp1基因整合进了家蚕的基因组,插入位点位于scaffold 868;(2)在RNA水平上,MaSp1的表达并不干扰其他3个基因(Fib-H, Fib-L and P25)的表达水平,并且外源基因MaSp1的表达水平略高于Fib-H;(3)在蛋白质水平上,MaSp1 蛋白的丰度大约占丝素总蛋白的2%,而且通过自身的LBS 区段和轻链蛋白(Fib-L)形成分子间二硫键,从而融入蚕丝蛋白复合体。
关键词:  转基因家蚕  蜘蛛牵引丝  蛋白质-蛋白质相互作用
DOI:
基金项目:Global COE资助。
Expression analysis of spiderdragline silk gene MaSp1 in the transgenic silkworm
HAN Leng,ZHANG Lei,NAKAGAKI Masao
(Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda 386-8567, Japan;Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda 386-8567, Japan; Sericultural Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230061)
Abstract:
To study the biological properties and structure of fibroin genes and proteins from transgenic silkworms, the transgenic silkworms were analyzed with inverse PCR, real-time PCR and co-IP methods. The results showed the following: (1) At the DNA level, MaSp1(major ampullate spidroin 1) was integrated into the B. mori chromosome at scaffold 868; (2 )At the RNA level, the transcription of MaSp1 did not affect the transcription of Fib-H, Fib-L or P25, and the transcription level of MaSp1 was higher than that of Fib-H; (3) At the protein level, MaSp1, the foreign silk protein, had low abundance among the total silk fibroin proteins (approximate 2%). Additionally, it had a protein-protein interaction with Fib-L through its LBS (L-chain-binding site) region through intermolecular disulfide bonds.
Key words:  transgenic silkworm  spider dragline silk  protein-protein interaction

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