昆虫学报2011，Vol.54Issue(2)：127-135,9.
棉铃虫围食膜肠粘蛋白基因HM72的克隆、表达及其蛋白组织定位
Cloning, expression and histolocalization of an intestinal mucin gene HM72 from Helicoverpa armigera (Hiibner) ( Lepidoptera: Noctuidae)
摘要
Abstract
Peritrophic membrane (PM) is an acellular semipermeable membrane secreted by midgut epithelial cells in most insects. The insect intestinal mucin is the most important component of PM. A fulllength cDNA clone HM72 encoding an intestinal mucin HM72 was identified by screening Helicoverpa armigera midgut cDNA expression library with the polyclonal antiserum against H. armigera PM proteins.The full-length cDNA of HM72 is 2 888 bp in length (GenBank accession no. HM017910), containing an open reading frame (ORF) of 2 469 bp, followed by an AT-rich untranslated region and a putative polyadenylation signal (AATTAA) located at 19 bp upstream of the polyA tail. The deduced protein sequences show that the HM72 is an acidic protein, and is synthesized as a preprotein of 823 amino acid residues with a 16-amino acid signal peptide. After removal of the signal peptide, the secreted HM72 protein is predicted to have a molecular weight of 84.2 kDa and isoelectric point of 3.63. The secreted HM72 has five chitin binding domains, one mucin domain and two glycine-aspartic acid-rich domain.HM72 was recombined into pET21b vector, and expressed by IPTG induction. Western blot analysis revealed that HM72 was more abundant in the exuviae, fecal pellets, peritrophic membrane and the midgut of H. armigera, and was secreted by cells from whole midgut. HM72 was not found in the larval fat bodies,integument, malpighian tubules, salivary glands, midgut fluid and hemolymph. This study provides a foundation for the research of bio-control associated novel genes of H. armigera and the perfection of the insect peritrophic membrane theory.关键词
棉铃虫/中肠/上皮细胞/围食膜/肠粘蛋白/cDNA表达文库/组织定位Key words
Helicoverpa armigera/ midgut/ epithelial cells/ peritrophic membrane/ intestinal mucin/cDNA expression library/ histolocalization分类
生物科学引用本文复制引用
刘小民,李杰,郭巍,徐大庆,张霞..棉铃虫围食膜肠粘蛋白基因HM72的克隆、表达及其蛋白组织定位[J].昆虫学报,2011,54(2):127-135,9.基金项目
国家重点基础研究发展规划("973"计划)项目(2009CB118902) （"973"计划）
国家自然科学基金项目(30971910) （30971910）
现代农业产业技术体系建设专项资金 （）
河北省科技支撑项目(09220302D) （09220302D）
