Differential enhancement of Cry2A versus Cry11A yields in Bacillus thuringiensis by use of the cry3A STAB mRNA sequence

FEMS Microbiol Lett. 1999 Dec 15;181(2):319-27. doi: 10.1111/j.1574-6968.1999.tb08862.x.

Abstract

Previously we demonstrated that the yield of Cry3A (70 kDa) can be increased as much as 10-fold when cry3A including its upstream STAB-SD mRNA stabilizing sequence is expressed in Bacillus thuringiensis under the control of cyt1A promoters. To determine whether the cyt1A promoters/STAB-SD combination (cyt1AP/STAB) has broader applicability, we used it to synthesize two other Cry endotoxins in the 70-kDa mass range, Cry2A and Cry11A. Combination of cyt1AP/STAB with orfs 2 and 3 of the cry2A operon yielded about 4. 4-fold the amount of Cry2A obtained with the wild-type cry2A operon. The yield of Cry11A obtained with a construct that contained the cyt1AP/STAB, cry11A and the 20-kDa protein gene was 1.3-fold the amount obtained with a construct similar to the wild-type operon. These results demonstrate that the cyt1AP/STAB combination can enhance synthesis of different Cry proteins significantly, but that the level of enhancement varies with the specific protein synthesized.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus thuringiensis / chemistry*
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism*
  • Bacterial Proteins / pharmacology
  • Base Sequence
  • Culicidae / drug effects
  • Larva / drug effects
  • Lepidoptera / drug effects
  • Molecular Sequence Data
  • RNA, Bacterial / genetics
  • RNA, Messenger / genetics
  • Regulatory Sequences, Nucleic Acid / genetics*

Substances

  • Bacterial Proteins
  • RNA, Bacterial
  • RNA, Messenger