C - Chemistry – Metallurgy – 07 – K
Patent
C - Chemistry, Metallurgy
07
K
530/13, 167/135,
C07K 14/285 (2006.01) A61K 39/102 (2006.01) C07K 16/12 (2006.01)
Patent
CA 1340712
The gene coding for Pasteurella haemolytica leukotoxin can be cloned in a plasmid expressed in Escherichia coli. The leukotoxin gene can be isolated from a clone bank of P. haemolytica. The clone bank is constructed by partial digestion of genomic DNA. The resultant 5 to 10 kilobase-pair fragments are ligated into plasmid vector pBR322. The resultant clones are screened for the production of P. haemolytica soluble antigens by a colony enzyme-linked immunosorbent assay blot method with a rabbit antiserum raised against the soluble antigens. The clones producing P. haemolytica soluble antigens are then analyzed for the production of the leukotoxin by a cytotoxicity assay with cells from a bovine leukemia-derived B-lymphocyte cell line as the target cells. Positive clones are identified, and subsequent restriction analysis of the recombinant plasmids shows the same insert DNA is cloned in the plasmid vector. The DNA sequence analysis of the insert DNA reveals regions coding for the leukotoxin.
616538
Lo Reggie Y. C.
Shewen Patricia C.
Strathdee Craig A.
Bereskin & Parr
University Of Guelph
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