CAJ | 학술논문

家蚕(Bombyx mori)的丝腺是丝蛋白合成分泌的场所,存在多种丝氨酸蛋白酶抑制剂.为探究家蚕丝蛋白的合成和保护机制,采用半定量RT-PCR的方法调查家蚕丝氨酸蛋白酶抑制剂基因serpin16在家蚕不同发育时期和5龄第5天幼虫各个组织器官中的表达特征,结果表明家蚕serpin16基因仅在4眠-5龄第6天的发育期表达,并且仅在丝腺中特异表达,其中在中部丝腺前区转录水平最高,而在中部丝腺中区和后区的转录水平较低;进一步构建pGEX-4T-1-serpin16原核表达载体,并转化至大肠杆菌(Eschevichia coli)BL21(DE3),经IPTG诱导获得融合蛋白, 纯化后得到单一的目的蛋白.家蚕serpin16基因在幼虫丝腺的特异表达模式提示其可能与家蚕的吐丝过程密切相关,推测该基因在维持丝腺稳定的泌丝环境中发挥重要作用.
가잠(Bombyx mori)적사선시사단백합성분비적장소,존재다충사안산단백매억제제.위탐구가잠사단백적합성화보호궤제,채용반정량RT-PCR적방법조사가잠사안산단백매억제제기인serpin16재가잠불동발육시기화5령제5천유충각개조직기관중적표체특정,결과표명가잠serpin16기인부재4면-5령제6천적발육기표체,병차부재사선중특이표체,기중재중부사선전구전록수평최고,이재중부사선중구화후구적전록수평교저;진일보구건pGEX-4T-1-serpin16원핵표체재체,병전화지대장간균(Eschevichia coli)BL21(DE3),경IPTG유도획득융합단백, 순화후득도단일적목적단백.가잠serpin16기인재유충사선적특이표체모식제시기가능여가잠적토사과정밀절상관,추측해기인재유지사선은정적비사배경중발휘중요작용.
Fibroin and sericin are synthesized in silk gland of the silkworm(Bombyx mori)where various serine protease inhibitors reside.In order to understand mechanism of silk protein synthesis and protection in silkworm,semi-quantitative RT-PCR was employed to investigate the expression patterns of silkworm serine protease inhibitor gene serpin16 at differ-ent developmental stages and in various tissues/organs of the 5th day larvae Of the 5th instar.The results showed that serpinl 6 was specifically expressed in silk gland from the 4th moulting to the 6th day of the 5th instar.Its transcriptional level in anterior region of the middle silk gland was the highest.The transcriptional levels in middle and posterior regions of the middle silk gland were relatively lower.Prokaryotic expression vector pGEX-4T-1-serpinl6 was further constructed and introduced into Escherichia coli BL21(DE3).Fusion protein was expressed successfully in the form of inclusion body through induction by IPTG.and further purified to obtain the single target protein.The exclusive expression pattern of silkworm serpin16 indicated that it is closely related with the process of silk spinning and may play an important role in maintaining the homeostasis for silk secretion in the silk gland.