Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli.

نویسندگان

  • H E Schnepf
  • H R Whiteley
چکیده

Sau 3A1 partial digestion fragments from Bacillus thuringiensis var. kurstaki HD-1 plasmid DNA were ligated into the BamHI site of the cloning vector pBR322 and transformed into Escherichia coli strain HB101. Colonies presumed to contain recombinant plasmids were screened for production of an antigen that would react with antibody made against B. thuringiensis crystals. One strain, ES12, was isolated by using this procedure. ES12 contains a plasmid of Mr 11 X 10(6) that has DNA sequence homology with pBR322 as well as with Mr 30 X 10(6) and Mr 47 X 10(6) plasmids of B. thuringiensis. It makes a protein antigen, detected by antibodies to crystal, which has the same electrophoretic mobility as the B. thuringiensis crystal protein. Protein extracts of ES12 are toxic to larvae of the tobacco hornworm Manduca sexta.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Molecular Cloning and Characterization of a Lipase from an Indigenous Bacillus pumilus

Cloning and sequencing of a lipase gene from an indigenous Bacillus pumilus, strain F3, revealed an open-reading frame of 648 nucleotides predicted to encode a protein of 215 residues. Sequence analysis showed that F3 lipase contained a signal peptide composed of 34 amino acids with an H domain of 18 residues. A tat-like motif was found in the signal peptide similar to some other Bacillus pumil...

متن کامل

Cloning and evaluation of gene expression and purification of gene encoding recombinant protein containing binding subunit of coli surface antigens CS1 and CS2 from Enterotoxigenic Escherichia coli

Background & Objective: Enterotoxigenic Escherichia coli (ETEC) is a major causative agent of diarrhea. Enterotoxins and the colonization factors (CFs) are major virulence factors in ETEC infections. The bacterium binds to the intestinal epithelial cell surface through colonization factors and produces enterotoxins that cause excessive fluid and electrolyte secretion in the lumen of the intesti...

متن کامل

MOLECULAR CLONING AND EVALUATION OF WILD PROMOTER IN EXPRESSION OF BACILLUS SPHAERICUS PHENYLALANINE DEHYDROGENASE GENE IN BACILLUS SUBTILIS CELLS

To evaluate the role of wild promoter of L-phenylalanine dehydrogenase (PheDH) gene, referred to as pdh, from Bacillus sphaericus in expression, cloning of pdh gene in Bacillus subtilis was performed. The whole pdh gene was cloned in pHY300PLK shuttle vector and amplified, construct (pHYDH) then transformed in B. subtilis ISW1214 and E. coli JM109. The pdh endogenous promoter presented no effec...

متن کامل

Cloning and optimization of phytase enzyme gene expression in Escherichia coli

Introduction Phytase is an enzyme that has the ability to break down phytic acid into myoinositol and mineral phosphate, and widely uses as an additive in animal foods. The aim of this study was to achieve a high level of bacterial phytase expression in PET26b expression host. Materials and Methods To generate the recombinant phytase enzyme, the target gene was introduced into the expression ...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 78 5  شماره 

صفحات  -

تاریخ انتشار 1981