人精液凝固蛋白SgI-52的表达与抗菌活性研究  

Expression of Human Semenogelin I-52 and Antibacterial Activity Investigation of Recombinant Peptide

作　　者：赵晖[1] 刘孝东[2] 高振华[2] 张建华[2] 李虹[3] 张云[1] 李文辉[1] 申吉泓[2] 

机构地区：[1]中国科学院昆明动物研究所动物模型与人类疾病机理重点实验室,云南昆明650223 [2]昆明医学院第一附属医院,云南昆明650032 [3]四川大学华西医院泌尿外科,四川成都610041

出　　处：《Zoological Research》2008年第2期139-144,共6页动物学研究（英文）

基　　金：国家自然科学基金(30670412);中国科学院"西部之光";云南省自然科学基金资助项目(2005PY01-23;2007C0006R)

摘　　要：以人精囊腺cDNA为模板,用嵌套式PCR技术扩增出编码成熟人精液凝固蛋白I(semenogelin I,SgI)第85–136位氨基酸(SgI-52)的核苷酸序列并将其插入载体pMAL-p2X中,成功构建的表达载体pMAL-p2X/SgI-52转化大肠杆菌DH5α后,重组融合蛋白诱导表达于大肠杆菌周质中。经第X因子剪切及超滤处理,得到表达的重组SgI-52。质谱分析结果证明重组SgI-52为目的肽。重组人SgI-52对大肠杆菌ATCC25922标准株以及耐氨苄标准株ML-35P的最低抑制浓度MIC分别为32.45以及46.30?μg/mL。我们的结果及相关研究提示在人精液液化过程中人精液凝固蛋白I的不同降解产物可能行使不同的生物学功能,值得进行深入研究。Nucleotide sequence coding for SgⅠ-52 with amino acid residues of 85-136 form mature human semenogelin Ⅰ was amplified by PCR technique from the cDNA of a human seminal vesicle. The obtained PCR products were inserted into vector pMAL-p2X. The constructed expression vector of pMAL-p2X/SgⅠ-52 was transformed into Escherichia coli DH5 α. Fusion protein was expressed in the periplasma of the E. coli DH5 α after IPTG inducement. Recombinant SgⅠ-52 was purified after factor X cleavage and a ultrafiltering process. MALDI-TOF- MS results indicated that the purified recombinant SgⅠ-52 was the target peptide. Recombinant SgⅠ-52 showed antibacterial activities on E. coli ATCC 25922 and E. coli ML-35P with MIC values of 32.45 and 46.30 μg/mL, respectively. Our results and other relevant works suggested that different human semenogelin Ⅰ degradation fragments during the liquefaction might have various biological functions and deserve to be investigated further.

关 键 词：人 精液凝固蛋白Ⅰ 表达 抗菌 

分 类 号：R392[医药卫生—免疫学]