Cloning Of Human Leukocyte Elastase Inhibition Factor Elafin Expression Vector And Its Sticky Protein Secretion Airway

PART ONE CLONING OF HUMAN NEW TYPE ANTIPROTEINASE PROTECTOR ELAFIN GENE CDNA AND THE CONSTRUCTION,IDENTIFICATION AND EXPRESSION OF RECOMBINED ADENOVIRUS VECTOR CONTAINING ELAFIN GENEObjective: To clone human new type antiproteinase protector elafin gene cDNA, to generate the adenoviral vector that expresses elafin gene for further study of elafin gene effects on the mucin secretion of pulmanory epithelial cell .Methods. Firstly, the total RNA was extracted from lung tissues,and elafin cDNA was gotten by means of RT-PCR. Secondly, the fragment was inserted into the multiple clone site of pAdTrack-CMV. The pAdTrack-CMV-elafin was analyzed by restriction endonuclease digestion, PCR, and sequence scan. Thirdly, the linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183. The candidate clone was further analyzed by restriction endonuclease digestion, PCR, and sequence scan. And then the recombined adenovirus plasmid was digested with Pac I and transfected into 293 cells for packaging and amplifying. Infection titer and rate were monitored by green fluorescent protein (GFP)...