[Construction and screening of TsF cDNA library specific for AchR].
OBJECTIVE
To construct the T suppress factor (TsF) cDNA library specific for AchR and screen out the positive clones of TsF.


METHODS
The full-length cDNA was synthesized by the reverse transcripting from the TsF PolyA+mRNA, which had been extracted from the AchR specific suppressor lines (ARSL). The cDNA library was obtained using Lambda gt11 vector. Finally, this cDNA library was screened by Western blotting using Analysis of this cDNA library showed that titer of the monoclonal antibody specific for TCR-achain.


RESULTS
recombinant bacteriophage was 1.4 x 10(7) pfu/ml, the rate of positive recombinant was 86.9%. The recombinant bacteriophage DNA were digested by EcoRI and electrophoresis analysis found that the main size of insertion was 0.8 to approximately 4.0 kb . As a result of screening, 27 positive clones were obtained. The positive recombinant bacteriophage DNA were digested by EcoRI, and electrophoresis analysis found that the size of insertion was 1 kb. Conclusion The results suggested that this cDNA library is available, the recombination phages can express TsF specific for AchR effectively and could be used to prepare the products of gene engineering of TsF.
Li
Wanyi
W
Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Li
Hong
H
Li
Mingyuan
M
Gong
Qimei
Q
Xiao
Liying
L
Jiang
Zhonghua
Z
D000818:Animals; D018076:DNA, Complementary / Q000096:biosynthesis*; D015723:Gene Library; D016147:Genes, Tumor Suppressor / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D010446:Peptide Fragments / Q000276:immunology; D011950:Receptors, Cholinergic / Q000235:genetics*; D011995:Recombination, Genetic; D050378:T-Lymphocytes, Regulatory / Q000378:metabolism*
D018076:DNA, Complementary; D010446:Peptide Fragments; D011950:Receptors, Cholinergic