Skip to main content
Log in

Construction and analysis of plasmids containing the Escherichia coli serB gene

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Plasmid pserB59-1 carries the E. coli serB gene on a 5.2 kb BamHI fragment cloned into the BamHI site of plasmid pBR322. The results of genetic and biochemical experiments established that a functional serB gene is contained in the fragment. The location of the serB gene within the insert was determined by restriction endonuclease analysis of plasmids derived from pserB59-1 that carry the Tn5 element at sites that inactivate the serB gene, and by deletions of segments of the 5.2 kb insert that either inactivate or do not inactivate the serB gene. A 38,000 Mr serB + polypeptide was detected when plasmid pserB59-1 was used as template in a minicell system, but not when the serB gene was inactivated by insertion of a Tn5 element.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ames BN (1966) Assay of inorganic phosphate, total phosphate and phosphatases. Methods Enzymol 8:115–118

    Google Scholar 

  • Berg DE (1977) Insertion and excision of the transposable kanamycin resistance determinant Tn5, in Bukhari, A.I., Shapiro, J.A. and Adhya, S.L. (Eds), DNA Insertion Elements, Plasmids and Episomes, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 205–212

    Google Scholar 

  • Berg DE, Davies J, Allet B, Rochaix J-D (1975) Transposition of R factor genes to bacteriophage λ. Proc Natl Acad Sci USA 72:3628–3632

    Google Scholar 

  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles II. A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Clewell DB (1972) Nature of Col El plasmid replication in Escherichia coli in the presence of chloramphenicol. J Bacteriol 110:667–676

    Google Scholar 

  • Guerry P, LeBlanc DJ, Falkow S (1973) General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol 116:1064–1066

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Analyt Biochem 114:193–197

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, p 68

    Google Scholar 

  • McKitrick JC, Pizer LI (1980) Regulation of phosphogycerate dehydrogenase levels and effect on serine synthesis in Escherichia coli K-12. J Bacteriol 141:235–245

    Google Scholar 

  • Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Pizer LI (1963) The pathway and control of serine biosynthesis in Escherichia coli. J Biol Chem 238:3934–3944

    Google Scholar 

  • Reeve J (1979) Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol 68:493–503

    Google Scholar 

  • Selker E, Brown K, Yanofsky C (1977) Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid Col E1. J Bacteriol 129:338–394

    Google Scholar 

  • Stauffer GV (1983) Regulation of serine, glycine, and one-carbon, biosynthesis. In: Herrmann KM, Somerville, RL (eds) Amino acids: Biosynthesis and genetic regulation. Addison-Wesley Publ. Co., Reading, MA

    Google Scholar 

  • Stauffer GV, Plamann MD, Stauffer LT (1981) Construction and expression of hybrid plasmids containing the Escherichia coli glyA gene. Gene 14:63–72

    Google Scholar 

  • Umbarger HE, Umbarger MA, Siu PML (1963) Biosynthesis of serine in Escherichia coli and Salmonella typhimurium J Bacteriol 85:1431–1439

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by G. O'Donovan

Rights and permissions

Reprints and permissions

About this article

Cite this article

Garnant, M.K., Stauffer, G.V. Construction and analysis of plasmids containing the Escherichia coli serB gene. Molec. Gen. Genet. 193, 72–75 (1984). https://doi.org/10.1007/BF00327416

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327416

Keywords

Navigation