1.Study On E-SLT,TNF\IL-8 And ET-1 In Lung Edema 2.Study On Apoptosis And P53 In Lung Cancer

CLONING,SEQUENCING AND POSITIONING OF THE ENCODING GENES OF β- LACTAMASES IN KLEBSIELLA PNEUMONIAEThe clinical resistant isolate 99-799 of Klebsiella pneumoniae was employed as the research object. And the encoding genes of β-lactamases (bla) in this strain was studied. K. Pneumoniae 99-799 was resistant to expanded-spectrum cephalorins,aztreonam,ciprofloxacin, someβ-lactamases inhibitors and gentamicin, but was susceptible to cefoxitin and imipenam. by This particular strain contained three plasmids (big, middle and small) that encoded the three β-lactamases with pIs of 5.4, 7.8 and 9.0, respectively.In the study, the 480, 748, 867 and 395-bp DNA fragment of β-lactamases encoding gene were amplified from K.pneumoniae 99-799 PCR with primers of bla, blaTEM, blaSHV and blaAmpC, respectively. Futhermore, the PCR products were purifed and cloned into PUCm-T vector and sequenced by sanger's dideoxy chain termination composition method. The results of sequence analysis showed that two of encoding genes were identified as TEM and SHV, in which the amino acid sequences were the same as that of TEM-1 and SHV-28, respectively; The other gene identified as the encoding AmpC-typeβ-lactamase was a 90% homologous to MIR-1, theβ-lactamase mediated by plasmid in K. pneumoniae was reported in 1990.In an attempt to isolate each plasmid and position the each resistant gene in K. Pneumoniae 99-799, the experiments of transformation, conjugation and PCR were carried out. The small and big plasmids, and the middle and big plasmids in K. Pneumoniae 99-799 were transferred into E.coli DH5α by transformation and conjugation, respectively. The results indicated that TEM-1 β-lactamase gene was on the small and big plasmids, and the genes encoded SHV-28β-lactamase and AmpC-typeβ-lactamase were found on the big and middle plasmids, respectivly. Qtherwise, the AmpC-typeβ-lactamase gene was also found in the chromosome of K.pneumoniae 99-799.In conclusion, the K.pneumoniae 99-799 produced threeβ-lactamases: TEM-1, SHV-28 and AmpC-typeβ-lactamase...