番木瓜PL和β-Gal基因双价反义表达载体的构建

Construction of PL and β-Gal antisense expression vector of papaya

在已报道的番木瓜果胶裂解酶(PL)和β-半乳糖苷酶(β-Gal)基因序列的基础上,设计特异引物,分别扩增保守区序列,将其分别反向插入植物表达载体pCAMB IA1301-35S-GUS-Nos的CaMV35S启动子和Nos终止子之间,构建反义表达载体pCP和pCG.然后用2种不同的方法将带有完整启动子和终止子的PL和-βGal基因引入pCAMB IA2301中,获得PL和β-Gal基因的双价植物反义表达载体pCPG,并对2种方法进行比较.

Pectate lyase and β-Galactosidase（β-Gal） are important softening enzyme.The transcriptions of PL and β-Gal gene were inhibited by antisense technique may put off papaya（Carica papaya L.） fruit softening.PL and β-Gal conserved domains were cloned using specific primers designed according to those published genes.The PL and β-Gal cDNA were inserted into between the CaMV35S promoter and Nos terminator of the expression vector pCAMBIA1301-35S-GUS-Nos with reverse orientation,respectively.So,two antisense expression vectors pCP and pCG were obtained.Then,PL and β-Gal antisense genes with 35S promoter and Nos terminator were inserted into expression vector pCAMBIA2301 together to generate double genes plant expression vector pCPG by different mode.