Datta A, Ghosh A K, Kundu S C
Department of Biotechnology, Indian Institute of Technology, 721302, Kharagpur, India.
Comp Biochem Physiol B Biochem Mol Biol. 2001 May;129(1):197-204. doi: 10.1016/s1096-4959(01)00377-3.
Fibroin gene expression during the larval developmental stages of the Saturniid silkworm, Antheraea mylitta, was analyzed. Northern blot analysis of larval silk gland total RNA using the fibroin gene as a probe showed that fibroin is expressed in the intermoult stages and repressed during the moulting stages. Abundance of fibroin transcripts gradually increased from the third to fifth intermoult stage, reaching a peak in the fifth intermoult. Transcripts declined during the early spinning stage. Western blot analysis of fibroin protein production with anti-fibroin antibody confirmed the differential fibroin expression, in accordance with fibroin mRNA synthesis. Dot blot hybridization of genomic DNA isolated from each larval developmental stage with the labelled fibroin gene showed that at the genomic level, the relative concentration of the fibroin gene was constant throughout the developmental stages. Our data confirm that fibroin gene expression in A. mylitta, like in B. mori, is transcriptionally controlled and shows differential temporal variations.
对柞蚕(Antheraea mylitta)幼虫发育阶段的丝心蛋白基因表达进行了分析。以丝心蛋白基因为探针,对幼虫丝腺总RNA进行Northern印迹分析,结果表明丝心蛋白在蜕皮间期表达,而在蜕皮期受到抑制。从第三次到第五次蜕皮间期,丝心蛋白转录本的丰度逐渐增加,在第五次蜕皮间期达到峰值。在吐丝早期转录本减少。用抗丝心蛋白抗体对丝心蛋白的产生进行Western印迹分析,证实了丝心蛋白的差异表达,这与丝心蛋白mRNA的合成一致。用标记的丝心蛋白基因对从每个幼虫发育阶段分离的基因组DNA进行斑点印迹杂交,结果表明在基因组水平上,丝心蛋白基因的相对浓度在整个发育阶段是恒定的。我们的数据证实,柞蚕中的丝心蛋白基因表达与家蚕一样,受到转录调控,并表现出不同的时间变化。