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produces protein epidermal growth factor urogastrone

DESCRIPTION OF VECTOR COMPONENT:
Name of vector: unknown
Intact vector size:
Type of vector: plasmid
Vector end: EcoRI
Vector end: SalI
Cloning sites:
Polylinker sites:
Host range: Escherichia coli
Features (with orientation and position when available):
marker(s): ampR
Cross references:

DNA: Synthetic

DESCRIPTION OF INSERT COMPONENT:
Genome: human
Gene symbol: EGF
Genomic copy number: unique
Gene name: epidermal growth factor
Contains complete coding sequence?: U
Chromosome: 4; Localization: 4 q25
Type of DNA: synthetic
Insert 5' end: EcoRI
Insert 3' end: SalI
Insert size (kb): 1.75
Cross references:

Insert lengths(kb): 1.75

Gene product: epidermal growth factor(urogastrone) [EGF]

Alleles: A1, B1, B2, A2

Target Gene: epidermal growth factor

Restriction digests of the clone give the following sizes (kb): EcoRI/SalI--3.3, 1.75; EcoRI--5.1; SalI--5.1; BamHI--4.9, 0.2.

Runaway vector contains a temperature-sensitive mutation in the copy-control gene. Maintained at a low copy number below 34C; replication increases 50-fold at temperatures greater than 34C. Growth at 37C is lethal.

The 1.75 kb EcoRI/SalI fragment contains a trp promoter/regulator and the EGF/urogastrone coding region. The complete coding sequence of the mature peptide, plus a "pro" sequence of 8 aa, is contained in a 0.2 kb XbaI/SalI fragment.

Directs the synthesis of a peptide of 61 aa, including an 8-residue "leader" sequence.

Expresses the urogastrone analog product on the order of 50 mg per O. D. liter as determined by SDS-PAGE. The microbially expressed product is biologically active.

Constructed from oligonucleotides selected to reflect the codon usage of Escherichia coli. Contains a HinfI site interrupting Asp3, an SphI site interrupting Cys31, and an NruI site after Tyr44.