| 胰岛激活蛋白B聚体的各亚基克隆及表达的初步研究 |
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| 引用本文： | 潘煦文.胰岛激活蛋白B聚体的各亚基克隆及表达的初步研究[J].新疆农业科学,2007,44(3):252-258. |
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| 作者姓名： | 潘煦文 |
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| 作者单位： | 新疆农业大学动物营养实验室,乌鲁木齐,830052 |
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| 基金项目： | 国家科技部重大基础研究前期研究专项资金 |
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| 摘 要： | 研究将百日咳杆菌用B-G培养基活化后再在改良的S-S培养基上液体培养,用CTAB-NaCl法提取百日咳杆菌的基因组DNA。经PCR扩增出胰岛激活蛋白B聚体的四个亚基基因,分别将其克隆于pGEX-6p-1载体上,并探讨了在大肠杆菌BL21(DE3)中可溶性表达IAP B聚体各亚基的可能性。通过控制诱导表达温度,S2,S3均可以实现可溶表达,以GST亲和层析得到的S2终产物0.4 mg/L培养物,S3为0.6 mg/L培养物。S4表达虽是可溶性的,但由于融合蛋白的等电点近于7,多步纯化易损失。S5的表达量太低,有待进一步分析。
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| 关 键 词： | 可溶性表达 胰岛激活蛋白 B聚体 GST融合蛋白 |
| 文章编号： | 1001-4330（2007）03-0252-07 |
| 收稿时间： | 2006-12-12 |
| 修稿时间： | 2006-12-12 |
Preliminary Study on Gene Clone and Expression About Four Subunits of Islet Activity Protein (IAP) B-oligomer |
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| PAN Xu-wen.Preliminary Study on Gene Clone and Expression About Four Subunits of Islet Activity Protein (IAP) B-oligomer[J].Xinjiang Agricultural Sciences,2007,44(3):252-258. |
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| Authors: | PAN Xu-wen |
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| Institution: | laboratory of Animal Nutrition, Xinjiang Agricultural University, Urumqi 830052, China |
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| Abstract: | In this research a Bordet-Gengou agar which contains sheep blood was used to culture the B.pertussis and then transfer single clone to a modified Stainer-Scholte broth to gain pure B.pertussis cells.The genome DNA of B.pertussis was extracted by using a cetryltrimethylammonium bromide(CTAB)-NaCl protocol.The four subunits of IAP B-oligoner were increased and were cloned on a recombinant pGEX-6p-1 vector by PCR,respectively.The results show that: S2 and S3 can be expressed in a soluble state and 0.4 mg/L(S2) and 0.6 mg/L(S3) were obtained,respectively after being purified by GST-affinity system;S4 subunit also can be expressed and purified but the yield is low because its being pI is close to 7;but S5 is hard to get the final product. |
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| Keywords: | soluble expression lAP B - oligomer GST - fusion protein |
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