中文摘要：

铜绿假单胞菌PA0057基因为1V类未知基因，通过NCBI比对，发现PA0057基因编码的蛋白与金属β-内酰胺酶同源性为96％，因此推测PA0057蛋白可能具有金属β-内酰胺酶活性．提取铜绿假单胞菌PAOI基因组DNA，利用设计的引物通过PCR方法扩增得到PA0057基因，将其克隆至克隆载体pGEM—T，并转化人大肠杆菌DH5α；而后将其克隆至表达载体pET28a中，转化入大肠杆菌BL21（DE3）中，IPTG诱导表达，并通过Ni—NTA亲和柱进行纯化，纯化后蛋白超滤浓缩得高浓度蛋白；通过药敏纸片检测PA0057蛋白活性．

英文摘要：

Through http..//www, pseudomonas, com/,we inference that the PA0057 protein may be a metal β-lactamase. Therefore, we intend to use PA0057 gene in Eseherichia coli expression and through a series of protein purification manner to expressed protein. Then we establish an analysis system that analyse the metal β- lactamase activity of PA0057 protein. The gene of P. aeruginosa PAOI extracted and the PA0057 gene was amplified by PCR with artificial primer. The amplified gene was recombined in cloning vector pGEM-T and transformed into E. coll. DH5a. The 0.9 kb DNA fragment containing PA0057 gene was detected. The PA0057 gene was cloned into expression vector pET28a,then transformed into E. coll. BL21（DE3）and afterward expressed by IPTG inducenment. The expression protein was purified by Ni-NTA His-Bind Resin. Then ,we used drug sensitive slips to test the activity of PA0057 protein.