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Enhanced expression of cro-beta-galactosidase fusion proteins under the control of the PR promoter of bacteriophage lambda.
EMBO J 1982; 1(10):1217-24EJ

Abstract

Hybrid plasmids carrying cro-lacZ gene fusions have been constructed by joining DNA segments carrying the PR promoter and the start of the cro gene of bacteriophage lambda to the lacZ gene fragment carried by plasmid pLG400 . Plasmids in which the translational reading frames of the cro and lacZ genes are joined in-register (type I) direct the synthesis of elevated levels of cro-beta-galactosidase fusion protein amounting to 30% of the total cellular protein, while plasmids in which the genes are fused out-of-register (type II) produce a low level of beta-galactosidase protein. Sequence rearrangements downstream of the cro initiator AUG were found to influence the efficiency of translation, and have been correlated with alterations in the RNA secondary structure of the ribosome-binding site. Plasmids which direct the synthesis of high levels of beta-galactosidase are conditionally lethal and can only be propagated when the PR promoter is repressed. Deletion of sequences downstream of the lacZ gene restored viability, indicating that this region of the plasmid encodes a function which inhibits the growth of the cells. The different applications of these plasmids for expression of cloned genes are discussed.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

6327257

Citation

Zabeau, M, and K K. Stanley. "Enhanced Expression of Cro-beta-galactosidase Fusion Proteins Under the Control of the PR Promoter of Bacteriophage Lambda." The EMBO Journal, vol. 1, no. 10, 1982, pp. 1217-24.
Zabeau M, Stanley KK. Enhanced expression of cro-beta-galactosidase fusion proteins under the control of the PR promoter of bacteriophage lambda. EMBO J. 1982;1(10):1217-24.
Zabeau, M., & Stanley, K. K. (1982). Enhanced expression of cro-beta-galactosidase fusion proteins under the control of the PR promoter of bacteriophage lambda. The EMBO Journal, 1(10), pp. 1217-24.
Zabeau M, Stanley KK. Enhanced Expression of Cro-beta-galactosidase Fusion Proteins Under the Control of the PR Promoter of Bacteriophage Lambda. EMBO J. 1982;1(10):1217-24. PubMed PMID: 6327257.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhanced expression of cro-beta-galactosidase fusion proteins under the control of the PR promoter of bacteriophage lambda. AU - Zabeau,M, AU - Stanley,K K, PY - 1982/1/1/pubmed PY - 1982/1/1/medline PY - 1982/1/1/entrez SP - 1217 EP - 24 JF - The EMBO journal JO - EMBO J. VL - 1 IS - 10 N2 - Hybrid plasmids carrying cro-lacZ gene fusions have been constructed by joining DNA segments carrying the PR promoter and the start of the cro gene of bacteriophage lambda to the lacZ gene fragment carried by plasmid pLG400 . Plasmids in which the translational reading frames of the cro and lacZ genes are joined in-register (type I) direct the synthesis of elevated levels of cro-beta-galactosidase fusion protein amounting to 30% of the total cellular protein, while plasmids in which the genes are fused out-of-register (type II) produce a low level of beta-galactosidase protein. Sequence rearrangements downstream of the cro initiator AUG were found to influence the efficiency of translation, and have been correlated with alterations in the RNA secondary structure of the ribosome-binding site. Plasmids which direct the synthesis of high levels of beta-galactosidase are conditionally lethal and can only be propagated when the PR promoter is repressed. Deletion of sequences downstream of the lacZ gene restored viability, indicating that this region of the plasmid encodes a function which inhibits the growth of the cells. The different applications of these plasmids for expression of cloned genes are discussed. SN - 0261-4189 UR - https://www.unboundmedicine.com/medline/citation/6327257/Enhanced_expression_of_cro_beta_galactosidase_fusion_proteins_under_the_control_of_the_PR_promoter_of_bacteriophage_lambda_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0261-4189&date=1982&volume=1&issue=10&spage=1217 DB - PRIME DP - Unbound Medicine ER -