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目的 获得大量完整的重组乙肝病毒（ Ｈ Ｂ Ｖ）ｐｒｅ Ｓ１ 多肽纯品，以利于ｐｒｅ Ｓ１ 功能与免疫学性质的研究。方法 比较不同表达载体及不同的培养、诱导和纯化条件，最大限度地使表达产物免受大肠杆菌蛋白酶的降解。结果 分析不同长度ｐｒｅ Ｓ１ 基因在两种不同载体，两种表型宿主菌中表达产物的状况，可看出在ｐｒｅ Ｓ１ 第５６ 和６４ 位氨基酸之间存在两个 Ｅ．ｃｏｌｉ 蛋白酶切割点。用ｐ Ｑｅ９载体和 Ｍ１５（ｐ Ｒ Ｅ Ｐ４） 宿主菌， Ｉ Ｐ Ｔ Ｇ 诱导１２ 小时以 Ｎｉ Ｎ Ｔ Ａ 琼脂糖柱在变性条件下纯化，能获得１０ ｍｇ／ Ｌ 以上具有良好免疫学活性的电泳纯ｐｒｅ Ｓ１ 蛋白。结论 缩短诱导时间、用一步法在变性条件下纯化表达产物可获得无明显降解的ｐｒｅ Ｓ１ 完整蛋白。
Objective To obtain a large number of complete recombinant hepatitis B virus (H B V) pre S1 polypeptide pure products in order to facilitate pre S1 function and immunological properties. Methods Different expression vectors and different culture, induction and purification conditions were compared to maximize the expression of the expressed product against degradation by E. coli proteases. Results Analysis of the expression of products of pre S1 gene of different lengths in two different vectors and two phenotypes showed that there are two E.coli between amino acids 56 and 64 of pre S1. Protease cleavage point. With pQe 9 vector and M15 (pR E P4) host bacteria, I P T G induced 12 hours to Ni NTA sepharose column in denaturing conditions to obtain 10 mg / L or more with good immunology Active electrophoresis pure pre S1 protein. Conclusion The induction time can be shortened, and the pre S1 intact protein without obvious degradation can be obtained by one-step purification of the expression product under denaturing conditions.