摘要
ＤＨＤＰＳ是赖氢酸合成通路中第一步的合成酶。将中国春小麦的ＤＨＤＰＳ基因质粒移植入ＲＤＡ８中，得到ＲＤＡ８／ｐＤＢ２６菌株，本研究通过细胞培养、层析提纯和结晶条件的探索，给出了一个较好的技术路线，并为开展义衍射分析蛋白结构创造了条件，通过该研究，对中国春小麦ＤＨＤＰＳ和野种大肠杆菌的ＤＨＤＰＳ差异有了一定的了解，并且对细胞培养的ＭＭ介质处理，ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ，Ｐｈｅｎｙｌ－Ｓｅｐｈａｒｏｓｅ和ＭｏｎｏＱ层析的方法给出具体实验条仲。酶活的检测和蛋白浓度测定都是采取高灵敏度的方法。结晶的ＳＣｒｅｅｎｉｎｇ条件对于二种ＤＨＤＰＳ有很大的差异。对野种大肠杆茵的ＤＨＤＰＳ，需要表面活性剂Ｎ－ｏｃｔｙｌ－Ｄ－ｇｌｕｃｏｐｙｒａｎｏｓｉｄｅ，ｐＨ１０．０～１０．５，对于中国春小麦一ＤＨＤＰＳ则未找到较好的表面活性剂，ｐＨ６．８～７．６。中国春小麦一ＤＨＤＰＳ晶体培养条件在以往文献中未见报导。
Dihydro dipicolinate synthase (DHDPS)is a first step on the lysine biosynthetic path-way. The plamids of the wild E. coli and Triticum aestivum containing DHDPS gene aresuccessfully expressed in RDA8 and RDA8/pDB26. The programs of purification to bothDHDPS from RDAS and from RDA8/pDB26 are different in few steps. Screening for crys-talline conditions is separatedly considered duing to the obviously thermal stability. The suit-able media for crystallization to both wild DHDPS and wheat DHDPS are in differences withPH, buffer media and salt concentration. We suggest that these differences hint some structurally characteristic behaviors,
