| 1. | The amount of the goal protein was evaluated by densitometric scanning . it indicated that the product of the vp2 gene was 14 % of total bacterial protein of dh5a
表达产物经ni - nta金属鏊合层析柱纯化后，得到了纯度较高的6his - vp2融合蛋白。 |
| 2. | The sds - page analysis revealed that the trka extracellular domains proteins were highly expressed and accumulated up to above 30 % of the total bacterial proteins after iptg induction
称a膜外域结构域重组蛋白表达量均占全菌蛋白的30 %以上。 |
| 3. | The amount of the goal protein was evaluated by densitometric scanning . it indicated that the 45ku product of the vp6 gene was 26 . 5 % of total bacterial protein of bl21
表达产物经ni - nta金属螯合层析柱纯化后，得到纯化的his - taggedvp6融合蛋白。 |
| 4. | Especially , the recombinant pet21a having 5 copy gene in e . coli bl21 ( de3 ) got high expression and expression level was up to 25 % of the total bacterial proteins
其中，含有5份串连基因的pet21a重组质粒在bl21 ( de3 )中的诱导表达量占总蛋白的25左右。 |
| 5. | 3 ' - utr of h pf4 c dna was deleted and tag was mutated to taataa . 2 after sds - page and densitometric scan analysis , the result show that expression level is 25 - 30 % of total bacterial proteins
山西医科大学2002届硕士学位论文一2dhsa pbv220 rhpf4经温控诱导表达后， sds page及凝胶密度扫描分析，表达产物占总国体蛋白的25 30 ，凝胶迁移特性与hpf4标准品相同。 |
| 6. | After washing with reagent , adopt the newest purification technology source30rpc , sds - page and densitometric scan analysis , the result show that expression level is 90 % of total bacterial proteins . after renaturation , ifnr , hgfa , hgfb , hpk5 were purified by akta purifier chromatogram instrument , sepharose fast flow , ssphacrayl series gel , selecting optimize condition . finally establish a kind of high efficient purification model of recombinant proteins produced in escherichia coli as inclusion bodies , purification product purity > 98 %
结论:总之，通过对发酵罐中重组工程菌各种培养因素的研究，建立了一种高密度、高表达发酵工艺体系，为重组蛋白的后续纯化提供了大量、稳定的原料供应;通过对不同目的蛋白的色谱行为的系统研究，建立了一种高效稳定、快速简洁、易于放大的包涵体重组蛋白分离纯化体系。 |
| 7. | In this study , we designed a pair of primers based on the sequence of the upstream and downstream of chicken il - 2 gene . about 600 bp chicken il - 2 cdna fragment was cloned from cona - stimulated chicken splenocytes by reverse transcription - polymerase chain reaction ( rt - pcr ) and was subcloned into puc18 vector . recombinant clone was demonstrated by restriction enzyme digestion and dna sequencing . next , we construct recombinant plasmid pproex ? t - il - 2 . the cdna of chicken il - 2 gene was subcloned into bamh i / hind iii sites of vector . the recombinant plasmid pproex ? t - il - 2 was transformed into e . coli dh5a and the bacteria was induced with iptg . it was demonstrated by sds - page and western blot that a 18kda protein which was equal to chicken il - 2 protein in molecular weight was expressed in e . coli dh5a . the expression level was up to 30 % of the total bacterial proteins . the purified protein was used to prepare the antibody against chicken il - 2 protein
经酶切鉴定及dna序列测定，该基因为鸡il - 2基因，其序列与sundick等报道的完全一致。在此基础上，我们把鸡il - 2基因亚克隆到大肠杆菌原核表达载体pproex ~ ( tm ) ht中，构建重组表达质粒并进行确证性序列测定，重组质粒测序结果表明将编码鸡il - 2成熟蛋白的基因正确地插入到原核表达载体pproex ~ ( tm ) ht的目的位点。重组质粒转化受体菌dh5后用iptg于37进行诱导培养， sds - page和westernblot分析显示，表达的鸡il - 2融合蛋白分子量约为18kda ，表达的融合蛋白经薄层扫描发现目的蛋白表达量约占菌体蛋白的30 。 |
| 8. | Sds - page analysis suggested that the bacteria containing the recombinant plasmid pet - 32a ( + ) - igf - i produced the fusion protein of 30kda as it was induced by iptg . consisting 10 % of the total bacterial proteins , and the pet - 30a ( + ) - igf - ii produced the fusion protein of 14kda , which consisting 35 % of total bacterial proteins . 5
Sds - page分析表明，重组质粒pet - 32a ( + ) - igf -在iptg诱导下表达分子量约30kda的融合蛋白，但其表达量不高，约为菌体总蛋白的10左右；重组质粒pet - 30a ( + ) - igf -在iptg诱导下表达分子量约14kda的重组蛋白，融合蛋白表达量约占菌体蛋白总量的35 。 |
| 9. | The recombinant expression plasmid was transformed into bl21 ( de3 ) and gene expression was induced by administration of iptg . expression was detected by sds - page and confirmed by western blot analysis , which shows that the yield of the target protein was approximately 30 % of the total bacterial protein
将这个表达载体转化入大肠杆菌表达菌株bl21 ( de3 )中， iptg诱导培养后，经sds - page和western杂交实验检测可知特异性表达带约在44kd的位置，其表达量大约占细菌总蛋白的30左右。 |
| 10. | Ptxb1 - hng and ptxb1 - m - insulin are expressed in e . coli successfully . after sds - page and densitometric scan analysis , the results show that the expression level of hng fusion protein is above 40 % and m - insulin fusion protein above 50 % of total bacterial proteins . western - blot result demonstrated m - insulin fusion protein had specific reaction with mouse anti human insulin antibody ( igg )
Pbv220 ? hng在大肠杆菌中未检测到表达，后两个克隆在大肠杆菌bl21 ( de3 )中获得高效表达， hng及m - insulin融合蛋白表达量分别占全菌蛋白的40及50左右；经western - blot鉴定m - insulin融合蛋白可以与小鼠抗人胰岛素单克隆抗体( igg )发生抗原抗体结合反应。 |
