title: 2A self-cleaving peptide-based multi-gene expression system in the silkworm Bombyx mori.
abstract: Fundamental and applied studies of silkworms have entered the functional genomics era. Here, we report a multi-gene expression system (MGES) based on 2A self-cleaving peptide (2A), which regulates the simultaneous expression and cleavage of multiple gene targets in the silk gland of transgenic silkworms. First, a glycine-serine-glycine spacer (GSG) was found to significantly improve the cleavage efficiency of 2A. Then, the cleavage efficiency of six types of 2As with GSG was analyzed. The shortest porcine teschovirus-1 2A (P2A-GSG) exhibited the highest cleavage efficiency in all insect cell lines that we tested. Next, P2A-GSG successfully cleaved the artificial human serum albumin (66 kDa) linked with human acidic fibroblast growth factor (20.2 kDa) fusion genes and vitellogenin receptor fragment (196 kD) of silkworm linked with EGFP fusion genes, importantly, vitellogenin receptor protein was secreted to the outside of cells. Furthermore, P2A-GSG successfully mediated the simultaneous expression and cleavage of a DsRed and EGFP fusion gene in silk glands and caused secretion into the cocoon of transgenic silkworms using our sericin1 expression system. We predicted that the MGES would be an efficient tool for gene function research and innovative research on various functional silk materials in medicine, cosmetics, and other biomedical areas.
authors.affiliation: State key laboratory of silkworm genome biology, Southwest University, Chongqing, China.|College of biology and technology, Southwest University, Chongqing, China.
authors.forename: Yuancheng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State key laboratory of silkworm genome biology, Southwest University, Chongqing, China.|College of biology and technology, Southwest University, Chongqing, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: State key laboratory of silkworm genome biology, Southwest University, Chongqing, China.
authors.forename: Riyuan
authors.initials: R
authors.lastname: Wang
authors.affiliation: State key laboratory of silkworm genome biology, Southwest University, Chongqing, China.|College of biology and technology, Southwest University, Chongqing, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.affiliation: State key laboratory of silkworm genome biology, Southwest University, Chongqing, China.|College of biology and technology, Southwest University, Chongqing, China.
authors.forename: Qingyou
authors.initials: Q
authors.lastname: Xia
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D010455:Peptides / Q000235:genetics* / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D047011:Silk / Q000235:genetics / Q000378:metabolism
chemical_list: D004527:Egg Proteins; D010455:Peptides; D011956:Receptors, Cell Surface; D047011:Silk; C030474:vitellogenin receptor; D005346:Fibroblast Growth Factors