| 黄司法，唐志敏，杨礼香.香蕉BBI 型蛋白酶抑制剂（MaBBI1）的诱导、变性和复性[J].广东农业科学,2016,43(9):44-50 |
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香蕉BBI 型蛋白酶抑制剂（MaBBI1）的诱导、变性和复性 |
| Induction，denaturation and renaturation of Musa acuminata Bowman-Birk type protease inhibitor（ MaBBI1） |
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| DOI：10.16768/j.issn.1004-874X.2016.09.007 |
| 中文关键词: 香蕉 Bowman-Birk 蛋白酶抑制剂（BBI） 诱导 变性 复性 |
| 英文关键词: Musa acuminata Bowman-Birk protease inhibitor（BBI） induction denaturation renaturation |
| 基金项目: |
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| 中文摘要: |
| BBI（Bowman-Birk protease inhibitor）是一种富含半胱氨酸的植物蛋白酶抑制剂，其抑制
的蛋白酶主要包括胰蛋白酶、糜蛋白酶和弹性蛋白酶等丝氨酸蛋白酶。以从香蕉（Musa acuminata L.
cv.Brazilian）根中提取的总RNA 逆转录成的cDNA 为模板，根据NCBI 公布的野生型马来西亚蕉（Musa
acuminata subsp. Malaccensis）BBI 基因（收录号：XM_009415512）的序列设计引物，克隆了1 个BBI 型
蛋白酶抑制剂基因（MaBBI1）。MaBBI1 基因大小为381 bp，编码126 个氨基酸，包括12 个半胱氨酸
（9.5%），将该基因构建到载体pGEX-6P-1 上转化大肠杆菌Rossetta 菌株进行重组蛋白GST-MaBBI1 的诱
导表达。在IPTG 诱导3 h 后，一个预测分子量为40.8953 ku 的GST-MaBBI1 融合蛋白被大量表达。IPTG 诱
导后的GST-MaBBI1 融合蛋白是不溶的并累积成包涵体，通过对包涵体进行变性和复性的方法得到可溶的
GST-MaBBI1 融合蛋白，为进一步在体外研究该蛋白的功能奠定基础。 |
| 英文摘要: |
| BBI（ Bowman-Birk protease inhibitor） is a kind of plant-derived protease inhibitor and rich in
cysteine，it can inhibit serine proteases，including trypsase，chymase，elastase and so on. A Bowman-Birk type
protease inhibitor gene（ MaBBI1） was cloned with the cDNA templates isolated and reversely transcribed from
total RNA of banana（ Musa acuminata L. cv.Brazilian） roots and the primers were designed according to the
sequence of wild Malaysian banana（ Musa acuminata subsp. Malaccensis） BBI gene（ GenBank: XM_009415512）
published in NCBI. The full length of MaBBI1 gene with 381 bp encoding 126 amino acids including 12 cysteines
（9.5%） was inserted into vector pGEX-6P-1 and transformed Escherichia coli Rossetta to induce the expression
of recombinant proteins GST-MaBBI1. A lot of GST-MaBBI1 fusion proteins with predicted molecular mass of
40.8953 ku were induced by IPTG at 3 h. However，the GST-MaBBI1 fusion proteins were insoluble after IPTG
induction and accumulating as inclusion bodies. To get the soluble GST-MaBBI1 fusion proteins，the inclusion
bodies were denatured and renatured. The soluble fusion proteins GST-MaBBI1 laid the foundation for its further
study in vitro. |
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