[Cloning and identification of human tissue-type plasminogen activator gene].
OBJECTIVE
To acquire the gene fragments coding for human tissue-type plasminogen activator (ht-PA).


METHODS
Total RNA was extracted from Bowes Melanoma cell line and reverse transcriptase-"touchdown" PCR was employed to amplify ht-PA cDNA that underwent digestion with the restriction enzymes and was subsequently cloned into vector pSP72 for transforming E.coli TG1. The cloned fragment was subjected to restriction mapping and sequence analysis for confirmation.


RESULTS
The restriction maps of the selected clones were consistent with those generated by computer-based analysis and was identical with that of reported ht-PA cDNA.


CONCLUSIONS
The gene fragment encoding ht-PA was successfully cloned, which lays a foundation for further study and artificial modification of this gene.
Li
Zhen-Lin
ZL
Department of Histology and Embryology, First Military Medical University, Guangzhou 510515, China.
An
Jin
J
D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D010959:Tissue Plasminogen Activator / Q000235:genetics*
D018076:DNA, Complementary; D010959:Tissue Plasminogen Activator