Cloning, sequence analysis, and overexpression of Escherichia coli folK, the gene coding for 7,8-dihydro-6-hydroxymethylpterin-pyrophosphokinase.

Abstract:

:The gene coding for the Escherichia coli enzyme 7,8-dihydro-6-hydroxymethylpterin-pyrophosphokinase has been cloned and sequenced. This gene, designated folK, codes for a protein of 159 amino acids, including an amino-terminal methionine. The protein was overexpressed in E. coli MC4100 by cloning the gene behind the lacUV5 promoter in a high-copy-number plasmid. The enzyme was purified to homogeneity. Amino-terminal analysis of the purified protein showed that the amino-terminal methionine had been removed. The compositional molecular mass (17,945 Da) was identical to the molecular mass determined by mass spectrometry. The enzyme was observed to have a large number of proline residues and migrated anomalously in sodium dodecyl sulfate-polyacrylamide gels, with an apparent molecular mass of 23,000 Da.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Talarico TL,Ray PH,Dev IK,Merrill BM,Dallas WS

doi

10.1128/jb.174.18.5971-5977.1992

keywords:

subject

Has Abstract

pub_date

1992-09-01 00:00:00

pages

5971-7

issue

18

eissn

0021-9193

issn

1098-5530

journal_volume

174

pub_type

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