CAJ | 학술논문

为了探讨家蚕Bombyx mori丝素蛋白重链信号肽序列在中部丝腺组织中是否具有功能活性,根据家蚕丝蛋白基因的启动子活性高、丝蛋白具有高效分泌的特性,构建了带有丝素重链基因fib-H信号肽的家蚕丝胶-1(ser-1)启动子(ser-HS),用ser-HS驱动DsRed 基因构建了分泌型瞬时表达载体pSK-SerHS-DsRed-polyA.转染细胞实验显示,该载体能在家蚕BmN细胞中瞬时表达DsRed.家蚕注射载体后,可在中部丝腺腔中检测到红色荧光,表明瞬时表达的DsRed已分泌到丝腺腔内.据此提出克隆的fib-H信号肽序列在家蚕中部丝腺组织中具有信号肽的功能.
위료탐토가잠Bombyx mori사소단백중련신호태서렬재중부사선조직중시부구유공능활성,근거가잠사단백기인적계동자활성고、사단백구유고효분비적특성,구건료대유사소중련기인fib-H신호태적가잠사효-1(ser-1)계동자(ser-HS),용ser-HS구동DsRed 기인구건료분비형순시표체재체pSK-SerHS-DsRed-polyA.전염세포실험현시,해재체능재가잠BmN세포중순시표체DsRed.가잠주사재체후,가재중부사선강중검측도홍색형광,표명순시표체적DsRed이분비도사선강내.거차제출극륭적fib-H신호태서렬재가잠중부사선조직중구유신호태적공능.
To verify whether the signal peptide sequence of fibroin H chain (fib-H) functionally possesses the activity in the middle silk gland, based on the characteristics of the strong promoter of the silk protein gene and the high level secretion of silk protein of the silkworm, Bombyx mori, we constructed the promoter ser-HS of sericin-1 gene (ser-1) with the signal peptide sequence of fib-H. Then, we constructed the transitorily secretory expression vector pSK-serHS-DsRed-polyA via driving the DsRed gene with the promoter ser-HS. We transfected pSK-serHS-DsRed-polyA into BmN cells by liposome. From the cells transfected with the vector, the red fluorescence could be detected, which verified that the expression vector could express DsRed in BmN cells transiently. Furthermore, when silkworm had been injected with the expression vector pSK-serHS-DsRed-polyA, red fluorescence could be observed in the lumen of the middle silk gland of silkworm. The results indicated that DsRed was expressed transiently and was secreted into the lumen of the middle silk gland. We concluded that the cloned signal peptide sequence of fib-H possesses the biological functions of the signal peptide in the middle silk gland.