Kloning Gen Kitinase Aeromonas caviae WS7b pada Bacillus subtilis 168

Abstract

Pests and diseases are the main problems in plant cultivation. Biological control is one of the control method of plant pathogen. Recently, chitinolytic bacteria has been used as biological control of plant pathogenic fungi widely. Biotechnology and genetic engineering have been successed determine the DNA sequences encoding chitinase (chiA) of A. caviae WS7b. This study to construct B. subtilis 168 transformant with promotor sigB-chiA fution. In this study, insertion of sigB-chiA fution to B.subtilis 168 chromosome was conducted by double crossover recombination into amyE locus. This research success to isolated plasmids pDL2 with inserts sigB-chiA from labolatory stock E. coli DH5α transformants and inserted to the amyE locus of B. subtilis 168 chromosome. B. subtilis 168 transformants capable to grow in medium containing chloramphenicol antibiotic, expressing chitinase on colloidal chitin agar, shown blue color with Gram staining to indicated that this transformant was B. subtilis 168.