关键词: 上海, 上海

Abstract: Arabidopsis WUS gene was correctly cloned, and the WUS gene constitutive plant expression vector and WUS-EGFP fusion gene inducible plant expression vector were successfully constructed. They were transformed into Agrobactrium tumefaciens EHA105. The calli of rubber tree were transformed by pCAMBIA2301-35S-WUS. The positive results of histochemical staining had been obtained. The process were the following. 35S-GUS fragment was cleavaged from PBI121 by double enzymes digestion and was cloned into pCAMBIA2301 to form pCAMBIA2301-35S-GUS middle plant expression vector. After extraction of the total RNA of Arabidopsis thaliana, WUS gene was cloned by RT-PCR method and linked into pEASY-T1 vector, then the vector was doubly digested to create WUS fragment, which thus was inserted into pCAMBIA2301-35S-GUS instead of GUS fragment and formed pCAMBIA2301-35S-WUS plant expression vector. On the other hand, EGFP gene was amplified from pCAMBIA2301-EGFP vector by PCR, then the fusion DNA fragment of WUS and EGFP which were amplified by gene splicing by overlap extension PCR was cloned into pER8 inducible plant expression vector. These two plant expression vectors were successfully transformed into Agrobactrium tumefaciens EHA105 by electroporation method. Double enzymes digestion, PCR and sequencing detection verified that the results were correct. The sequences of the cloned WUS gene and WUS-EGFP fusion gene were the same as NCBI′s, it showed that these two vectors had been correctly constructed. Afterwards, friable embryogenic calli of rubber tree were transformed by the vector pCAMBIA2301-35S-WUS, and GUS staining showed that transformed calli had been obtained. These studies had provided a good base for further researching on the function of WUS gene in somatic embryogenesis and organogenesis of calli in Hevea brasiliensis.