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Structure characteristics and expression profiles of Bombyx mori α1 (IV) collagen gene, a temperature‐sensitive lethality‐related gene
Author(s) -
Ji M.M.,
Lu Y.J.,
Gan L.P.,
Niu Y.S.,
Sima Y.H.,
Xu S.Q.
Publication year - 2010
Publication title -
journal of applied entomology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 60
eISSN - 1439-0418
pISSN - 0931-2048
DOI - 10.1111/j.1439-0418.2009.01471.x
Subject(s) - biology , exon , bombyx mori , microbiology and biotechnology , gene , complementary dna , bombycidae , genetics , gene expression , sequence analysis , peptide sequence
The type IV collagen is a heterotrimer of two α1 (IV) and one α2 (IV) chains, which are encoded by emb ‐9 and let ‐2 genes in Caenorhabditis elegans, respectively, and the amino acid mutations in Gly‐X‐Y repeat region can cause temperature‐sensitive lethality during late embryogenesis. Here, we introduced a way to quickly and effectively do research on Bmlet ‐2 ( BmColIVα 1) gene from Bombyx mori , a central model for Lepidoptera. BmColIVα 1 gene is 6.583 kbp in full length, containing four exons, and its cDNA is 3.847 kbp, which contains an ORF for a protein of 819 amino acids with a molecular weight of 82.1 kDa and an isoelectric point of 6.43. The predicted protein sequence has closer relationship with α1 (IV) collagen chain of other species, and contains two conserved regions of collagen and nine PFAM domains of collagen triple helix repeat regions. Moreover, transmembrane region analysis and subcellular localization analysis reveal that this protein belongs to extracellular secreted protein. The experimental expression profile demonstrated that the expression level of BmColIVα 1 was obviously higher in head and fat body on day 3 of the fifth instar. The mutations of BmColIVα 1 gene located in exon 2 and exon 3 were identified in temperature‐sensitive lethal strain sch and wild stain Dazao . Seven mutations – namely, 242(T/A), 279(T/A), 556(T/C), 675(C/T), 1343(T/C), 1463(A/G), and 1574(T/C) – were novel, and five of them would modify the encoded amino acids, namely, 81(L/Q), 186(S/P), 448(L/S), 488(Q/R), and 525(V/A). The results of molecular modelling showed that the five amino acids mutations were located in four regions of 3‐helixs repetitive element sequence of ColIVα1 protein. BmColIVα 1 protein is a member of the collagen family, and its gene might be a temperature‐sensitive lethality‐related gene.