| 猪传染性胸膜肺炎放线杆菌apxIA基因在毕赤酵母中的表达与分析 |
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| 引用本文： | 黄琦,谢芝勋,庞耀珊,童桂香,文艳玲,谢志勤,邓显文,刘加波,谢丽基.猪传染性胸膜肺炎放线杆菌apxIA基因在毕赤酵母中的表达与分析[J].畜牧与兽医,2008,40(4):19-23. |
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| 作者姓名： | 黄琦 谢芝勋 庞耀珊 童桂香 文艳玲 谢志勤 邓显文 刘加波 谢丽基 |
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| 作者单位： | 1. 广西大学动物科技学院预防兽医系,广西,南宁,530001
2. 广西兽医研究所生物技术实验室,广西,南宁,530001 |
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| 基金项目： | 广西科技攻关项目资助(桂科攻0537008-3B);广西区水产畜牧局科研计划(桂渔牧科061910). |
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| 摘 要： | 构建猪传染性胸膜肺炎放线杆菌(APP)apxIA基因的真核表达载体pPICZαA/apxI,并在毕赤酵母GS115中进行表达。SDS-PAGE显示仅在浓缩40倍的上清中检测得到表达产物,同时经RT-PCR可检测到重组酵母中编码目的基因成熟肽的mRNA,经分析发现目的序列AT含量高达62%,其中含49个毕赤酵母稀有密码子,占15.4%。证实ApxIA在毕赤酵母中为低水平表达。
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| 关 键 词： | 胸膜肺炎放线杆菌 apxIA基因 毕赤酵母 表达 |
| 文章编号： | 0529-5130(2008)04-0019-05 |
| 修稿时间： | 2007年10月23 |
Expression and analysis of apxIA gene from Actinobacillus pleuropneumoniae in Pichia pastoris |
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| HUANG Qi,XIE Zhi-xun,PANG Yao-shan,TONG Gui-xiang,WEN Yan-ling,XIE Zhi-qing,DENG Xian-wen,LIU Jia-bo,XIE Li-ji.Expression and analysis of apxIA gene from Actinobacillus pleuropneumoniae in Pichia pastoris[J].Animal Husbandry & Veterinary Medicine,2008,40(4):19-23. |
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| Authors: | HUANG Qi XIE Zhi-xun PANG Yao-shan TONG Gui-xiang WEN Yan-ling XIE Zhi-qing DENG Xian-wen LIU Jia-bo XIE Li-ji |
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| Abstract: | A recombinant eukaryotic expression vector of apxIA gene from Actinobacillus pleuropneumoniae (App), named pPICZαA/apxI, was constructed and the apxIA gene was expressed in Pichia pastoris. SDS-PAGE analysis revealed that the recombinant ApxIA protein could be tested in the concentrated supernatant, and recognized by positive serum against APP serotype 1 using western-blot. The mRNA coding the ApxIA could be tested by RT-PCR in the recombinant yeast. In the interest sequence, the AT% was up to 62% and there were 49 rare codons of P.pastoris which was 15.4% of the total amount of amino acids. The results confirmed that ApxIA was expressed at a lower level in P.pastoris. |
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| Keywords: | Actinobacillus pleuropneumoniae apxIA gene Pichia pastoris expression |
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