Molecular cloning and expression of bovine kappa-casein in Escherichia coli

Journal Article · Fri Jan 01 04:00:00 EST 1988 · J. Dairy Sci.; (United States)

DOI:https://doi.org/10.3168/jds.S0022-0302(88)79521-1· OSTI ID:5244243

Kang, Y C; Richardson, T

A cDNA library was constructed using poly(A)/sup +/RNA from bovine mammary gland. This cDNA library of 6000 clones was screened employing colony hybridization using /sup 32/P-labelled oligonucleotide probes and restriction endonuclease mapping. The cDNA from the selected plasmid, pKR76, was sequenced using the dideoxy-chain termination method. The cDNA insert of pKR76 carries the full-length sequence, which codes for mature kappa-casein protein. The amino acid sequence deduced from the cDNA sequence fits the published amino acid sequence with three exceptions; the reported pyroglutamic acid at position 1, tyrosine at position 35, and aspartic acid at position 81 are, respectively, a glutamine, a histidine, and an asparagine in the clone containing pKR76. The MspI-, NlaIV-cleaved fragment (630 base pair) from the kappa-casein cDNA insert has been subcloned into expression vectors pUC18 and pKK233-2, which contain a lac promoter and a trc promoter, respectively. Escherichia coli cells carrying the recombinant expression plasmids were shown to produce kappa-casein protein having the expected mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and being recognized by specific antibodies raised against natural bovine kappa-casein.

• Research Organization: Univ. of Wisconsin, Madison
• OSTI ID: 5244243
• Journal Information: J. Dairy Sci.; (United States), Journal Name: J. Dairy Sci.; (United States) Vol. 71:1; ISSN JDSCA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
550401 -- Genetics-- Tracer Techniques
553005 -- Agriculture & Food Technology-- Animal Husbandry-- (1987-)
59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
AMINO ACID SEQUENCE
ANIMALS
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CASEIN
CATTLE
CLONING
COWS
DAYS LIVING RADIOISOTOPES
DNA
DNA SEQUENCING
DNA-ASE
DNA-CLONING
DOMESTIC ANIMALS
ENDONUCLEASES
ENZYMES
ESCHERICHIA COLI
ESTERASES
GENETIC MAPPING
GLANDS
HYBRIDIZATION
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
MAMMALS
MAMMARY GLANDS
MAPPING
MICROORGANISMS
MOLECULAR BIOLOGY
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHOSPHODIESTERASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
RECOMBINANT DNA
RUMINANTS
STRUCTURAL CHEMICAL ANALYSIS
TRACER TECHNIQUES
VERTEBRATES