Discovering genes responsible for silk synthesis in Bombyx mori by piggyBac-based random insertional mutagenesis
Discovering genes responsible for silk synthesis in Bombyx mori by piggyBac-based random insertional mutagenesis
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通过基于piggyBac的随机插入诱变发现负责家蚕丝合成的基因
DOI：
10.1111/1744-7917.12595
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发表时间：
2019
期刊：
影响因子：
4
通讯作者：
Huang Li Hua
中科院分区：
文献类型：
--
作者：
Feng Xing Bao;Zheng Zi Wen;Zhang Xian;Gu Jun;Feng Qi Li;Huang Li Hua
家蚕突变体是转基因育种和基因发现的宝贵资源。基于PiggyBac的随机插入突变已广泛用于基因功能研究。为了发现与蚕丝合成相关的基因，以家蚕为材料构建了piggyBac-随机插入文库，并对异常茧突变体进行了筛选。通过这种方法，鉴定了一个“薄茧”突变体。与野生型相比，突变体茧层变薄，后部丝腺变短。在突变体的PSG中，所有三种丝素蛋白基因（包括Fib ‐H、Fib‐ LandP 25）的信使RNA（mRNA）水平均显著下调。在水通道蛋白（AQP）、缺乏蛋白样纵向通道（Lola）、谷氨酰氨基肽酶样（GluAP）和Loc 101744460中发现了4个piggyBac插入位点。所有四个基因的mRNA水平显着改变的丝腺突变体。特别是，mRNA量ofAQP，一个负责调节渗透压的基因，显着下降之前立即纺纱阶段在前丝腺的突变体。本研究对“薄茧”突变体中缺失的基因进行了鉴定，为进一步了解家蚕的产丝规律和转基因育种提供了有用的信息。
Silkworm mutants are valuable resources for both transgenic breeding and gene discovery. PiggyBac‐based random insertional mutagenesis has been widely used in gene functional studies. In order to discover genes involved in silk synthesis, a piggyBac‐based random insertional library was constructed usingBombyx mori, and the mutants with abnormal cocoon were particularly screened. By this means, a “thin cocoon” mutant was identified. This mutant revealed thinner cocoon shell and shorter posterior silk gland (PSG) compared with the wild type. The messenger RNA (mRNA) levels of all the three fibroin genes, includingFib‐H,Fib‐LandP25, were significantly down‐regulated in the PSG of mutants. Four piggyBac insertion sites were identified inAquaporin(AQP),Longitudinals lacking protein‐like(Lola),Glutamyl aminopeptidase‐like(GluAP) andLoc101744460. The mRNA levels of all the four genes were significantly altered in the silk gland of mutants. In particular, the mRNA amount ofAQP, a gene responsible for the regulation of osmotic pressure, decreased dramatically immediately prior to the spinning stage in the anterior silk gland of mutants. The identification of the genes disrupted in the “thin cocoon” mutant in this study provided useful information for understanding silk production and transgenic breeding of silkworms in the future.