PURPOSE: To provide sheep LFA-3 protein which has high affinity to human T-cells, thus being useful as a diagnostic reagent for detecting T-cells, by culturing the cells transformed with a vector including the DNA sequence coding the sheep LFA-3 and collecting the product after cultivation.
CONSTITUTION: RNA is extracted from sheep leukocytes with guanidine thiocyanate and the extract is treated with oligo (dT) cellulose column to isolate mRNA and cDNA is synthesized using it as a template and further prepare cDNA library. Then cloning is effected using it as a probe to collect the gene encoding sheep LFA-3 protein. The gene is multiplied by the PCR(polymerase- chain reaction) process and treated with a restriction enzyme to link to a suitable vector and introduced into host cells such as Escherichia coli and transformed. The transformed cells are cultured in a medium to enable high- efficient production of the objective sheep LFA-3 protein in high efficiency.
OOHARA TAKAAKI
NIWA HIDEO
KOSAKATA FUMIO
FUKUCHI TAKESHI
EDAMURA TADAHIRO
SUMIYA TORU
