使用偏爱密码子大幅度提高苯丙氨酸脱氨酶在食品级乳酸乳球菌中的表达_陈星.pdf

DOI :10.13345/j.cjb.2006.02.003 22 2 　　　　　 Vol.22 　No.2 2006 3 ChineseJournal of Biotechnology March　2006 　 High_level Expression of Phenylalanine Ammonia_lyase in Lactococcus Lactis Via Synthesized Sequence Based on Bias Codons 1, 2 1,2 1 1 1 1 1 1 1 ＊ 陈　星 , 高　斌 , 贾兴元 , 苏　畅 , 吕月平 , 王战勇 , 范新萍 , 肖　白 , 刘敬忠 1, 2 1, 2 1 1 1 1 CHEN Xing , GAO Bin , JIA Xing_Y an , SU Chang , L Y e_Ping , WANG Zhan_Yong , FAN 1 1 1 ＊ Xin_Ping , XIAO Bai and LIU Jing_Zhong 1 , 　100020 2 2003 , 　100054 1 Bejiing Chaoyang HospitalAffiliate of Capital University of MedicalSciences, Beijing 100020, China 2 Master Graduateof Capital University of MedicalSciences, Bejiing 100054, China nat 　　为获得苯丙氨酸脱氨酶 (PAL)在食品级乳酸乳球菌中的高效表达, 将欧芹pal cDNA (pal )及根据乳酸乳球菌 art 偏爱密 子设计人工合成的pal 基因 (pal )重组并转化到两种乳酸乳球菌NICE 诱导表达系统中, 测定基因工程菌表达 PAL 酶的量及活性, 对比分析密 子偏爱性对乳酸乳球菌表达外源蛋白的影响。 结果表明:在两种乳酸乳球菌NICE 表达系 统中, 使用偏爱密 子均可显著提高PAL 酶的表达效率, 使NZ9000 pNZ8048 表达系统表达量提高22.23 倍, NZ3900 pNZ8149 系统提高35.90 倍。此研究获得了安全高效表达PAL, 可用于治疗苯丙酮尿症的基因工程菌。 　 苯丙氨酸脱氨酶, 乳酸乳球菌, NICE 表达系统, 密 子 　Q786　　　A　　1000-3061 (2006)02-0187-04 Abstract　To constr ct a safer and more efficient gene engineering LactococcusLactis for expressing phenylalaine ammonia lyase (PAL)which will be benefit for PKU therapy, pal cDNA of Parsly and synthesized seq ence based on Lactococcus Lactis bias codons were recombined into two Lacto