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KMID : 0366219890240020441
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Korean Journal of Hematology
1989 Volume.24 No. 2 p.441 ~ p.454
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A Study of Bioactive Substance Preparation From Human Lymphocyte by Genetic Engineering-Technique and Its Anti-cancer Effect
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이수영/Soo Young Lee
박원상/김준호/이기평/김주성/최진/Won Sang Park/Joon Ho Kim/Ki Pyung Lee/Choo Seung Kim/Jin Choi
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Abstract
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In order to investigate the immunomodulatory and tumoricidal effects of bioactive
substances, CDNAS have been prepared from poly(A)+ mRNAs isolated
from human lymphocytes stimulated with cancer promoter, TPA (12-o-tetradecanoyl
phorbol 13-acetate) and PHA-p. Poly(A)+ mRNAs purified from total
cellular RNAs of the TPA-PHA stimulated human lymphocytes have been fractionated
by sucrose del)sixty gradient centrifugation. Of 32 mRNA fractions protein synthesis has
been shown in 13 fractions by in vitro translation.
The cDNAS have been synthesized on the oligo(dT)-tailed pBR 322-SV40 primer by
the use of fraction number 14 and 16 mRNAs as templates, followed by annealing with
the oligo(dT)-tailed pBR 322-SV40 linker and replacement of RNA strands by DNA. E.
coli K-12(HB 101) have been transfected by the synthesized CDNA and screened by
dot-blot hybridization using 32P-mRNAs as probes. Sonicated a supernatants and
SDS-extracts from the E. coli transformants have been characterized by slab-gel
electrophoresis, 3H-thymidine uptake test of human lymphocytes, antiviral
activity assay, interleukin-2 activity assay, and tumoricidal activity assay using human
uterine cervix carcinoma cell line, MEl80 as target cell.
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