PMID: 6308625Jul 1, 1983

Open reading frame expression vectors: a general method for antigen production in Escherichia coli using protein fusions to beta-galactosidase

Proceedings of the National Academy of Sciences of the United States of America
G M WeinstockM Zweig

Abstract

We have developed an Escherichia coli plasmid vector for the identification and expression of foreign DNA segments that are open reading frames (ORFs). The 5' end of ompF, an E. coli gene encoding an abundant outer membrane protein, is used to provide a strong, regulated promoter, translation initiation site, and signal sequence for export from the cytoplasm. This sequence is coupled to the lacZ gene of E. coli so that expression of beta-galactosidase requires ompF transcription and translation signals. However, this hybrid gene is LacZ- because lacZ is out of frame with respect to ompF. Restriction enzyme recognition sites are located between ompF and lacZ to allow convenient insertion of DNA fragments. If an insert is an ORF of the correct length, ompF and lacZ become realigned in frame, resulting in a LacZ+ gene that produces a tribrid protein with the translation product of the insert sandwiched between OmpF and beta-galactosidase. The LacZ+ phenotype thus identifies clones containing an expressed ORF. To demonstrate the vector's utility we inserted a fragment from the herpes virus thymidine kinase gene and used the resulting tribrid protein to raise antibodies that precipitate thymidine kinase from herpes virus-infected ce...Continue Reading

References

May 1, 1988·Plant Molecular Biology·S J SandlerP Dunsmuir
Apr 1, 1986·Molecular & General Genetics : MGG·W SpevakH Ruis
Jan 6, 1992·Biochimica Et Biophysica Acta·V G Wilson, J Ludes-Meyers
Jan 1, 1983·Biotechnology Advances·J D Friesen, G An
Jan 30, 2002·Gene·Irene T RombelStephen Albert Johnston
Aug 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·D O Clegg, D E Koshland
Aug 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·L S ThomashowM F Thomashow
Oct 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·T KiyokawaM Yoshida
Jun 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·M YamadaT Grodzicker
Aug 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·C S Hoffman, A Wright
Apr 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·E ElfassiJ L Dienstag
Jun 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·P S MeissnerM L Berman
Oct 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·D B LimW K Maas
Nov 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J D Liu, J S Parkinson
Apr 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·X RuanN K Peters
Jan 1, 1985·Biotechnology & Genetic Engineering Reviews·J D BrookA L Meredith
Aug 1, 1989·AIDS Research and Human Retroviruses·N KomiyamaM Yoshida
Nov 12, 1984·Nucleic Acids Research·J Shlomai, A Zadok
Apr 11, 1988·Nucleic Acids Research·D WallsF Gannon
Dec 1, 1986·Journal of Clinical Pathology·A F Wright
Feb 1, 1986·Journal of Medical Genetics·D J ShawP S Harper
Apr 6, 2007·Scoliosis·Theodoros B Grivas, Olga D Savvidou
Mar 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·C A Davis, S Benzer
Jan 1, 1990·Archives of Virology·J A LenstraB A van der Zeijst

Citations

Feb 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A M Maxam, W Gilbert
Nov 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·U RütherB Müller-Hill
Mar 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·M J WagnerW C Summers
Jul 10, 1981·Nucleic Acids Research·S Anderson
Jan 30, 1982·Virology·P V McGuirtG B Elion
Feb 7, 1980·Nature·D E BüchelB Müller-Hill
Sep 5, 1981·Journal of Molecular Biology·M N Hall, T J Silhavy
Nov 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·M R GrayM Rosbash
Mar 1, 1981·Cell·T HoriiH Ogawa
Jan 1, 1981·Annual Review of Genetics·M N Hall, T J Silhavy
Mar 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·K Talmadge, W Gilbert

Related Concepts

Thymidine Kinase
GLB1
Alkalescens-Dispar Group
expression vector
Immune Sera
Promoter
Herpes virus antigen
Antigens, Bacterial
Antigens
Genes, Bacterial

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Sexual Dimorphism in Neurodegeneration

There exist sex differences in neurodevelopmental and neurodegenerative disorders. For instance, multiple sclerosis is more common in women, whereas Parkinson’s disease is more common in men. Here is the latest research on sexual dimorphism in neurodegeneration

HLA Genetic Variation

HLA genetic variation has been found to confer risk for a wide variety of diseases. Identifying these associations and understanding their molecular mechanisms is ongoing and holds promise for the development of therapeutics. Find the latest research on HLA genetic variation here.

Super-resolution Microscopy

Super-resolution microscopy is the term commonly given to fluorescence microscopy techniques with resolutions that are not limited by the diffraction of light. Here are the latest discoveries pertaining to super-resolution microscopy.

Genetic Screens in iPSC-derived Brain Cells

Genetic screening is a critical tool that can be employed to define and understand gene function and interaction. This feed focuses on genetic screens conducted using induced pluripotent stem cell (iPSC)-derived brain cells.

Brain Lower Grade Glioma

Low grade gliomas in the brain form from oligodendrocytes and astrocytes and are the slowest-growing glioma in adults. Discover the latest research on these brain tumors here.

CD4/CD8 Signaling

Cluster of differentiation 4 and 8 (CD8 and CD8) are glycoproteins founds on the surface of immune cells. Here is the latest research on their role in cell signaling pathways.

Alignment-free Sequence Analysis Tools

Alignment-free sequence analyses have been applied to problems ranging from whole-genome phylogeny to the classification of protein families, identification of horizontally transferred genes, and detection of recombined sequences. Here is the latest research.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.