Studies on Sex-lethal Gene of Bombyx Mori and Bombyx Mandarina
|Course||Special Economic Animal Feeding|
|Keywords||Silkworm Bombyx mandarina Sex-determining gene|
The gender of the silkworm are the female Shaped gametes decision mechanism, there has been no substantial progress in the feminizing the Fem genes of research. Drosophila Sxl gene is a master switch gene of sex determination. In this paper, a new discovery of Bombyx mori Bm-Sxl spatial and temporal differences in gene expression and gender differences of Bm-Sxl gene in the role of the silkworm sex determination may. Bm-Sxl gene according to the published sequence (Teruyuki Niimi, et al. 2006) designed specific primers Bm-Sxl gene was amplified by RT-PCR technology to analyze. Bombyx mori Bm-Sxl gene in Bombyx mori fifth instar gut expression; fifth instar larval and pupal stage ovarian expression, no significant period differences in expression levels; fat body and blood, the mature period Bm-Sxl The abundance of gene expression was significantly lower than in other periods. Bombyx mori Bm-Sxl gene 0.5 h of eggs in Virgin moth eggs, and birth not expressed, but in the age of the embryo in the egg 30 h transcriptional activity start time similar to Drosophila Sxl gene activation, suggesting that Bombyx mori Bm-Sxl gene may and sex determination. Bm-Sxl gene analysis showed that expression of gender differences, male and female egg stage expression differences Bm-Sxl gene is specifically expressed in the female egg, and further speculated Bm-Sxl gene may be related to sex determination. Wild Lepidoptera and after the artificial domestication feeding thousands of years silkworm genetic variation may exist. Two contains the complete ORF was cloned from the wild silkworm eggs silkworm Sxl gene in other Lepidoptera genetic differences Bmand-Sxl gene fragments (Bmand-Sxl-1 1 218 bp, Bmand- Sxl-2 1 279 bp), prokaryotic expression. Of Bombyx mandarina the Bmand-Sxl gene cloning sequencing, silkworm corresponding sequence than found, only 11 and 7 nucleotide differences homology of 99.1% and 99.4%. Bombyx mandarina Bmand-Sxl-1 gene fragment was cloned in the pET-28a () BamH I and EcoR I sites between, and speculate that the expression of fusion protein comprises 341 amino acid residues in its structure contains two conserved RNA recognition motif RRM. SMS protein molecular weight online toolkit to calculate the molecular size of the fusion protein to 37.77 k. The recombinant plasmid-containing BL21 total bacterial protein electrophoresis and Western blot hybridization can be detected is consistent with the theoretical molecular weight protein band. Therefore, wild the silkworm Bmand-Sxl-1 gene with His6 sequence the successful fusion expression in E. coli, In vitro studies Bmand-Sxl gene function conditions.