The Isolation Of BeCRN1 From Bambusa Emeiensis'Viridiflavus'and Study On Genetic Transformation

In order to study the function of BeCRN1, the homologous Gene of AtCRN1, and establish the genetic transformation system of Bambusa emeiensis'Viridiflavus', this species was selected as raw materials to do the following research:BeCRN1 was isolated from the leaves of Bambusa emeiensis'Viridiflavus'by the technology of rapid amplification of cDNA ends (RACE); Bioinformatics analysis of BeCRN1 was made;The plant expression vector pCHF3-BeCRN1 was constructed and introduced; The antibiotic sensitivity test of Bambusa emeiensis'Viridiflavus'callus was done; Transient expression detection of the GUS gene was done by histochemical method on Bambusa emeiensis'Viridiflavus'callus. The results indicated that:(1)BeCRN1 has a total length of 1646bp with an open reading frame of 1473bp and encodes a protein of 491aa. It has a 5 'UTR with the length of 45bp and a 3' UTR with the length of 128 bp.(2)NCBI Blast Search indicated that BeCRN1 has the highest homology with a protein from Oryza sativa(EEC80574.1)in the amino acid sequence. The identity and similarity between them are 85% and 91% respectively. The identity and similarity with CRN1 from Arabidopsis thaliana (NP₁96884.1) are 59% and 75% respectively.(3)The plant expression vector pCHF3-BeCRN1 was constructed and subse- quently introduced into Agrobacterium tumefaciens strain GV3101 via the freeze- thaw transformation method.(4) Transient expression of GUS detection indicated that the callus of Bambusa emeiensis'Viridiflavus'probably has backgroundβ-glucuronidase activity.(5)The antibiotic sensitivity test showed that the callus of Bambusa emeiensis'Viridiflavus'was not sensitive to the kanamycin, PPT and hygromycin.