[DNA family shuffling of tissue-type plasminogen activator genes].
OBJECTIVE
To probe the possibility of screening tPA with higher activity through DNA shuffling.


METHODS
Human, rat and rhesus monkey tPA cDNAs were used as the templates of "DNA family shuffling". The diverse tPA library was expressed in CHO cells, then the screening was carried out.


RESULTS
Two shuffled clones, t9 and t17 were selected. The activity of tPA expressed by t9 clone was 4-fold higher than the activity of human tPA. tPA expressed by t17 clone was found having a 88-amino-acid deletion, but still showed the same activity as human tPA. DNA sequencing demonstrated that the sequences of t9 and t17 were mainly derived from human and monkey tPA cDNAs.


CONCLUSIONS
Above work has laid the foundation for further experiments in the screening for higher activity of tPA.
Huang
Qing-sheng
QS
Institute of Microbiology and Epidemiology, AMMS, Beijing 100071, China.
Xu
Jing
J
Xu
Jun-jie
JJ
Tong
Yi-gang
YG
Wang
Hai-tao
HT
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D032621:DNA Shuffling; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008253:Macaca mulatta; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D010959:Tissue Plasminogen Activator