Differential enhancement of Cry2A versus Cry11A yields in Bacillus thuringiensis by use of the cry3A STAB mRNA sequence.
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.
Park
H
H
Department of Entomology, University of California, Riverside, CA 92521, USA.
Bideshi
D K
DK
Johnson
J J
JJ
Federici
B A
BA
D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D009033:Culicidae / Q000187:drug effects; D007814:Larva / Q000187:drug effects; D007915:Lepidoptera / Q000187:drug effects; D008969:Molecular Sequence Data; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*
D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger