Isolate and purify the target protein . prepare 50ml e . coli expressing the interesting proteins and harvest the cells and determinate the solubility of the target protein . completely solubilize inclusion bodies with 8m urea 溶解包涵体，将包涵体溶解物上ni一taslurry ，利用该融合蛋白表达时带有的his一tag与ni +的亲和作用分离、纯化表达蛋白。
Section hi : purify & refold recombinant hpk - 5 protein was efficiently expressed in e . coli jm109 as inclusion bodies . after bacteria were smashed by ultrasound , te buffer , 1 % ttiton x - 100 and 2 m urea was used to efficiently extract inclusion bodies 用含sm尿素溶液洗涤包涵体， 8000r / min转速下分离包涵体，能最大限度去除杂蛋白，同时不会降低目的蛋白的损失。
After the expression form analysis , the insoluble recombinant proteins was purified by destraction and abstersion of inclusion bodies . to study the abstersion condition of the inclusion bodies , we adopted ultrasound crushing and freezing - melting methods 采用超声加洗涤液破碎菌体；离心加冻融分离纯化融合蛋白，研究不同的超声次数和冻融对包涵体洗涤效果的影响。
It is the foundation of therapy of hepatitis b . the gene of scfv was cloned into the expression vector pet32a ( + ) and subsequently expressed in escherichia coll . the expressed scfv fused with the 109 aa trx - tag thioredoxin protein was above 30 % total cell protein in the shaking flask cultures 融合硫氧还蛋白的单链抗体在大肠杆菌bl21 ( de3 )中以包涵体的形式得到高效表达，单链抗体表达量达占30的总蛋白以上。
The inclusion bodies of recombinant protein were purified with washing buffer consisting of various urea ( 2mol / l and 4mol / l ) for several times , and then dissolve the fusion protein in the denature buffer using 8mol / l urea as denaturant 用含有2mol l和4mol l尿素的包涵体洗涤液洗涤包涵体，在37条件下，洗去了大部分菌体蛋白及其它核酸物质。用8mol l尿素作为变性剂溶解包涵体，包涵体在8mol l尿素中的溶解性非常好。
Although some pga precursors formed inclusion bodies in the cytoplasm of dh5a / pkkfpga , the amount only made up 5 - 10 % of all the afpga peptides , this explained why the expression level of afpga in dh5 / pkkfpga was a little lower than that in dh5 / psmlfpga 虽然青霉素g酰化酶在菌株dh5 pkkfpga中形成包涵体，但其量只占总酶的5 - 10 ，解释了为什么菌株dh5 pkkfpga的表达量虽然比菌株dh5 pkkfpga低，但差距不大。
After washing with reagent ( 50mmol / l ph8 . 0 tris - hcl , 100mmol / l nacl , 0 . 5mmol / l edta , 2mol / l carbamide , 0 . 2 % triton x - 100 , 0 . 2 % doc ) , ultrasound crushing ( 200 w , 150 times , 3 seconds per time , spacing 3 seconds ) and freezing - melting methods , we got hng fusion protein and m - insulin fusion protein with purity of above 80 % . 4 . radioimmunoassay result shows that the radioimmune activity of of m - insulin fusion protein is 0 . 5 unit per litre bacterial liquid 3 .表达产物初步纯化仁gly ”一flumanin融合蛋白和人胰岛素突变体融合蛋白包涵体的洗涤方案为: lj ]体沉淀溶于含zmol ， / l尿索洗涤液，超声( 200w 、巧o次、 3秒/次、间隔3秒) ， 4 、 i000orpm离心15min后沉淀用洗涤液充分重悬，一2 ( )冻存过夜，次日融化后4 、 10000rpm离心15min ，取上清，即得到初步纯化的融合蛋白，可去除土要的杂蛋白，融合蛋白溶解于上清液中，纯度达到80 %以_ 1 : 。
Sectionii : to research and establish a kind of high efficient purification model of recombinant proteins produced in escherichia coli as inclusion bodies . to raise separate efficiency and biological active efficiency is especially important in the purification model of recombinant proteins . in accordance with these questions , we raise inclusion bodies purity before the purification as far as possible , and select the suitable chromatogram technology 研究并建立一种包涵体中高效分离纯化目的蛋白的优化模式重组蛋白分离纯化工艺中分离效率的提高、生物活性效率的提高及工艺的稳定性是尤其重要的，针对这些问题，我们在层析纯化前尽山西医科大学生物化学与分子生物学2003届硕士学位论文可能地提高包涵体纯度，并选择合适层析方法的同时合理的组合色谱分离单元。
The steady dead generation and time that was caused by the isolated virus was certain by chicken embryo which was inoculated on seven or nine days . the histopathological changs of the infectious stunting syndrom were studied by the way of ordinary paraffin section and he dying . the experimental result were as follows : the test proved that the changes of the chicken embryo were different in different stage . the chicken embryo dead in a week after it inoculated . the body was dropsy and hemorrhage . dead before it hatched out , the embyo body were dropsy , pale and slime . the liver was yellow and swolled , gallbladder ( vesica fellea ) was filled with bile . bursa and glandula thymus analosis . the kindey dropsy . bowel lamina were humble , dilatation . gas and yellow foam were filled the bowel . histopathological changes were that , in early stage , obvious changes of liver and kindey were dropsy , hemorrhage and necrosis . two types eosinophilic intranuclear inclusion bodies including large round and little granular were present in cells of the above organs . the obvious changes of bursa were dropsy , adverse folliiculated growth and little lymphocytes proliferating , 19 - 21 days chicken embryo , one or two big empty vacuoles were prensent in cells of liver and kindey . the number of the folliculi was growing , the vacuoles between cells were larger 胆囊充盈、其内充满稀薄的胆汁；法氏囊、胸腺萎缩，肠道扩张、肠壁菲薄、内充满气体及黄色泡沫状物；肾脏肿大。病理组织学变化方面，早期肝脏、肾脏、肠主要以出血、水肿和坏死为主，且肝细胞核及肾小管的上皮细胞核内均发现有核内包涵体，包涵体呈嗜酸性，为大型圆形包涵体或不规则的颗粒状；法氏囊则以水肿、滤泡发育不良、小型淋巴细胞数量增多为主。 19 21日龄鸡胚肝细胞、肾小管上皮细胞的胞浆内出现1 2各大的空泡，法氏囊滤泡数目增多细胞间有较大空隙。
