Expression of the crylAcl Gene Under the Control of the Native or the $\alpha$-Amylase Promoters in an Acrystalliferous Bacillus thuringiensis Strain

  • Roh, Jong-Yul (Department of Plant Protection, Huazhong Agricultural University) ;
  • Lee, In-Hee (Department of Plant Protection, Huazhong Agricultural University) ;
  • Li, Jian-Hong (Department of Plant Protection, Huazhong Agricultural University) ;
  • Li, Ming-Shun (Graduate School of Agricultural Biotechnology, Seoul National University) ;
  • Kim, Ho-San (Graduate School of Agricultural Biotechnology, Seoul National University) ;
  • Je, Yeon-Ho (Graduate School of Agricultural Biotechnology, Seoul National University) ;
  • Boo, Kyung-Saeng (Graduate School of Agricultural Biotechnology, Seoul National University)
  • Published : 2000.12.01

Abstract

Expression of the crylAcl gene of an acrystalliferous Bacillus thuringiensis strain under the control of the native or $\alpha$-amylase gene promoter was investigated. The crylAcl gene was cloned in a B. thuringiensis - E. coli shutle vector, pHT3101, undder the control of either the native promoter (pProAc) or the $\alpha$-amylase promoter from Bacillus subtilis (pAmyAc). These two recombinant plasmids were successfully expressed in B. thuringiensis subsp. kurstaki Cry B. The first transformant (ProAc/CB), harboring pProAc, expressed an about 130 kDa protein begining 24 hr after inoculations just as in the case of the wild type of B. thuringiensis subsp. kurstaki HD-73. The second pAmyAc-transformant (AmyAc/CB) began to express the gene just 6 hr after inoculation, but Western analysis showed that the activity of the $\alpha$-amylase promoter was relatively weaker than that of the native promoter. As expected, their toxicity against Plutella xylostella larvae was dependent on the amount of Cry1Acl protein expressed.

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