Banana Bunchy Top Virus Chinese Zhangzhou Isolate Dna4 Coding Region Of Gene Cloning And Prokaryotic Expression

Banana is one of the five important fruit in the world. It plays an very important role in the international trades. Banana Bundchy top disease (BBTD) is the most serious disease of banana which infected by Banana Bunchy top virus (BBTV). Molecular biology research of BBTD began more than 20 years ago.BBTV is an single strand RNA (ssDNA) virus. The genome of BBTV has six components, BBTV DNAK 2^ 3> 4, 5x 6 respectively. BBTV DNA 4 encodes a putive trasmember move protein, but its function in whole plant is unkown.The aim of this experiment is cloning of encoding rgeion of BBTV DNA 4, expression the gene in E:coli and preparation its specific serum which may provided an improtant proof in testing of the ruction of protein encoded by BBTV DNA 4 in the whole plant.The coding region of banana bunchy top virus Chinese Zhangzhou isolate (BBTV-ZZ) DNA 4 was cloned by PCR. The result of sequence analysis shows that it is 351 nucleotides long and putatively encodes a protein of 116 amino acids.The taget gene was constructed into a series of prokaryotic expression vectors. The prokaryotic fusion expression vector pTYB-B4 for BBTV DNA 4 open reading region was constructed by cloning in pTYB2. Result from SDS-PAGE showed that the 69kD protein encoded by the fusion gene was expressed correctly in E.coli ER2566.The fusion protein was obtained from SDS-PAGE and antiserum against the protein was raised in rabbit. Western blot been analysis showed 14.5kD target protein expressed in transgenic tobacco which Southern blot has been analyzed.This expriment has provided a important evidence that the protein enconded by BBTV-ZZ DNA 4 might have function of viral movement protein in intact plant.Inadditional, effectors that effect the foreign genes expression high efficiently in E.coil were also discussed.