CAJ | 학술논문

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.
이용반향PCR、정량PCR이급면역공침정등방법분석연구전기인가잠적사소단백급기인적결구화성질.결과표명:(1)재 DNA 수평상,MaSp1기인정합진료가잠적기인조,삽입위점위우 scaffold 868;(2)재 RNA수평상,MaSp1적표체병불간우기타3개기인(Fib-H, Fib-L and P25)적표체수평,병차외원기인MaSp1적표체수평략고우Fib-H;(3)재단백질수평상,MaSp1단백적봉도대약점사소총단백적2%,이차통과자신적LBS 구단화경련단백(Fib-L)형성분자간이류건,종이융입잠사단백복합체.
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 re-sults 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 lev-el, 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.