摘要
应用重组苜蓿银纹夜蛾核型多角体病毒(AcMNPV)为基因转移载体,以绿色荧光蛋白基因(EGFP)为报告基因,研究不同截短的丝素重链基因启动子(-2127/+23、-925/+23和-238/+23)驱动报告基因在家蚕5龄幼虫组织中的表达情况。结果发现,通过病毒感染介导,3种长度丝素重链基因启动子驱动的报告基因在脂肪体和血细胞中存在异位表达,从个体体表可直接观察到明显的绿色荧光;3种长度的启动子都能在后部丝腺中高效起始EGFP的转录,产生绿色荧光;长度为0.9kb和0.2kb的启动子0.9KH、0.2KH在中部丝腺能起始EGFP转录产生绿色荧光,而2.1kb的启动子(2.1KH)能起始EGFP转录,但未能检测到绿色荧光;3种长度的启动子也能在Sf9培养细胞中起始EGFP基因表达。上述结果表明,丝素重链基因启动子没有严格的组织特异性。
In order to explore the regulation mechanism of fibroin heavy chain gene in silkworm （Bombyx mori）, the recombinant AcMNPV was used as the transfer vector and green fluorescent protein （EGFP） gene was used as the reporter gene to investigate the expression profiles of the reporter gene in silkworm tissues of the 5th instar larvae driven by differently truncated promoters of fibroin heavy chain gene （FibH） （-2 127/＋23, -925/＋23 and -238/＋23）. The results showed that there was ectopic expression of EGFP in fat body and hemocyte which was driven by three FibH promoters of various lengths. Green fluorescence could be observed directly on silkworm cuticle under fluorescence microscope. The three promoters of various lengths could initiate transcription of EGFP efficiently in posterior silk gland and produced green fluorescent light. Promoters of 0.9KH and 0.2KH could initiate transcription of EGFP in middle silk gland and yielded green fluorescent light. That of 2.1KH could initiate transcription of EGFP but no green fluerescent light was detected. All three promoters could also drive ectopic expression of EGFP in Sf9 cultured cells. The above results revealed that the promoter of fibroin heavy chain gene does not have strict tissue specificity.
出处
《蚕业科学》
CAS
CSCD
北大核心
2010年第3期391-399,共9页
ACTA SERICOLOGICA SINICA
基金
国家重点基础研究发展计划“973”项目(No.2005CB-121000)
教育部长江学者和创新团队发展计划项目(No.IRT0750)
