| 猪胃蛋白酶原A在毕赤酵母中的表达 |
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| 引用本文： | 刘西梅,安立国,乔宪凤,唐青海,华文君. 猪胃蛋白酶原A在毕赤酵母中的表达[J]. 江汉大学学报(自然科学版), 2008, 36(3): 59-63 |
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| 作者姓名： | 刘西梅 安立国 乔宪凤 唐青海 华文君 |
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| 作者单位： | 1. 湖北省农业科学院畜牧兽医研究所,动物胚胎工程及分子育种湖北省重点实验室,武汉,430064
2. 西北农林科技大学,动物科技学院,陕西,杨凌,712100 |
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| 基金项目： | 湖北省科技厅重点攻关项目，动物胚胎及分子育种湖北省重点实验室开放课题 |
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| 摘 要： | 利用已有的猪胃蛋白酶原AcDNA,构建毕赤酵母pPIC3.5K-pA胞内表达载体.经纤维蛋白消化试验和干酪素琼脂平板法筛选,获得两株高表达的重组子,研究它们在不同pH值、不同诱导时间、不同起始浓度和不同甲醇诱导浓度等条件下表达水平的高低.结果表明:在pH值为7、诱导72h、起始诱导浓度的OD600为2、甲醇诱导浓度为1%时,重组胃蛋白酶活力最高,达到6.46U/mL.
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| 关 键 词： | 猪胃蛋白酶原A 毕赤酵母 表达 |
Expression and Condition Optimization of Pepsinogen A in Pichia Pastoris |
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| LIU Xi-mei,AN Li-guo,QIAO Xian-feng,TANG Qing-hai,HUA Wen-jun. Expression and Condition Optimization of Pepsinogen A in Pichia Pastoris[J]. Journal of Jianghan University(Natural Sciences), 2008, 36(3): 59-63 |
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| Authors: | LIU Xi-mei AN Li-guo QIAO Xian-feng TANG Qing-hai HUA Wen-jun |
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| Affiliation: | LIU Xi-mei1,AN Li-guo2,QIAO Xian-feng1,TANG Qing-hai2,HUA Wen-jun1, |
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| Abstract: | Based on the cloning and sequence analysis of pepsinogen A from the Hubei swine,constructed plasmid of pPIC3.5K-pA in Pichia pastoris.There were screened two over-expression of recombinants by the experiment of hemaleucin digestion and the method of casein agar plate,and studied the effect of different pH,different induction time,different induction concentration and methanol induction concentration on the expression yield.The results indicated that,the activity of recombinant pepsin was able to reach 6.46 U/mL,under the conditions that the pH value was 7,induct 72 h,the induction concentration OD600 was 2,the methanol induction concentration was 1%. |
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| Keywords: | pepsinogen A Pichia pastoris expression |
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