DETAILED DESCRIPTION The present invention is directed to the means and methods of producting human antithrombin III via recombinant DNA technology, including (1) the discovery and identity of the entire DNA sequence of the mature protein as well as its signal polypeptide, and the 3'- and 5'-flanking regions thereof; (2) the construction of cloning and expression vehicles comprising said DNA sequence, enabling the amplification, and thence, expression of the mature human antithrombin III protein, as well as Met, fusion, or signal N-terminus conjugates thereof; and (3) viable cell cultures, genetically altered by virtue of their harboring such vehicles and capable of producing human antithrombin III polypeptide.
Further, this invention provides human antithrombin III in physical state distinct from its existence in, or isolation from, a natural environment or source, it by virtue of its method of preparation herein, being essentially free of usual endogenous proteins and other native materials or substances.
This invention is directed to the recombinant DNA production of human antithrombin III in all of its aspects, and is not to be construed as limited to any specific details described herein and embraced within the compass of this invention.
For example, the term "mature" as used herein connotes human ATIII as well as methionyl as a first amino acid, present by virtue of the ATG translational codon in the expression vector construction hereof.
DESCRIPTION OF PREFERRED EMBODIMENTS The work described herein was performed employing, inter alia, the microorganism E.
coli K-12 strain 294 (end A, thi.
sup.
-, hsr.
sup.
-, .
sub.
k hsm.
sup.
+), as described in British Patent Application Publication No.
2055382A.
This strain has been deposited with the American Type Culture Collection, ATCC Accession No.
31446, on Oct.
28, 1978.
However, various other microbial strains are useful, including known E.
coli strains such as E.
coli B, E.
coli .
times.
1776 (ATCC No.
31537, deposited July 3, 1979) and E.
coli W3110 (F
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Gene Therapy 
