Cloning And Vector Construction And Transformation By RNAi Of CBL In Soybean

Soybean contains high proteins, well quality, and excellent nutritive. Soybean proteins is determined an exquisite article on plant proteins.Cystanthionine (3-lyase consents to be the second critical enzyme in the path of methionine biosynthesis, and also be the critical enzyme in cysteine metabolic pathway. CBL plays an important role in metabolic pathway of sulfur-containing amino acid.To identify the characteristic on CBL, the research took some studys on several respects:Firstly, full-length cDNA GsCBL was cloned from the wild soybean (Glycine soja) Sequence analysis showed that, this gene has two copies in the wild soybean, and located in chromosome of NO.03 and NO.19. There is no diversity obviously; both of them have 13 exons and 12 introns. Tissue expression analysis showed that GsCBL was lowest in stem, followed by leaves and roots, and flowers were the highest in these four tissues. Then, GsCBL at different time of seeds grows was determination. When the seeds grow for 50 days, GsCBL has the highest expression. It is nearly twice time than flowers.The research analyzed the feature of GsCBL, and construction the GsCBL over-expression vector. Such information could be an importance tools to improve the sulfur-containing amino acids in soybean.RNA interference (RNAi) is a post-transcriptional gene-silencing by double-strand RNA. RNAi builds up a reliable and rapid research skill on reverse genetics. After interference the target genes, functions can be identification by the changed phenotype of transgene plant.The research designs a couple of idioprimer on GsCBL to construction RNA interference vector by Gateway technology. It will be a favorite tool for characterization of GsCBL genes on soybean genome.Basis on genetic transformation of soybean, the research took some simple test at soybean germination. This research found, sterilization by chlorine about 9-12 hour could be the best time to the seeds; and determination that hygromycin could take good effect on 1-5 mg/1. GsCBL-i was transferred into soybean.Two plants was got and showed different phenomenon to compare with cultivar.