| 1. | Of the differentially expressed clones , 104 clones were highly expressed in the ovary at stage iii , the remaining clones were highly expressed in the ovary at stage ii
结果共获得2倍以上差异表达克隆167个。其中期高表达克隆104个，期高表达克隆63个。 |
| 2. | Conclusion constructed the high - level expression clone of echistatin in e . coli . the expression of recombinant protein is higher , it make the further study of echistatin feasible
结论成功构建了echistatin的原核高效表达克隆，表达量高于现有国内外研究水平，为echistatin功能及相关疾病的研究奠定了基础。 |
| 3. | Chapter 3 sequencing cdna fragments of genes related to the ovary development of the mitten crab ( eriocheir japonica sinensis ) partial clones were sequenced which were selected from the differentially expressed clones screened by cdna macroarray analysis
3中华绒螯蟹卵巢发育相关基因cdna片段的序列测定从cdna微阵列筛选的差异表达克隆中选取部分克隆进行测序。 |
| 4. | Conclusion we have constructed the expression plasmid pbv220 - hpf4 successfully . 3 ' - utr of h pf4 c dna was deleted and tag was mutated to taataa by pcr . after sds - page and densitometric scan analysis , the expression level of r hpf4 is 25 - 30 %
结论本研究运用pcr定点突变技术，完全去除了hpf4cdna基因3 ”端utrat富含区：改用大肠杆菌强串联终止密码子taaaataa ，成功构建高效表达克隆pbv220 rhpp4 。 |
| 5. | In the first part , the focus is to find the receptor molecules directly by screening two cdna libraries with a recombinant construct prpap or as an alternative , to find an enriched area in the embryo brains and construct libraries from this brain region and perform the expression cloning as above
方法： （ ）以融合蛋白prpap通过瞬时表达克隆法筛选两个cdna文库，或者通过与胚胎脑的结合实验筛选有丰富结合蛋白的脑区，以图构建cdna文库并进行表达克隆的筛选。 |
| 6. | After sds - page and densitometric scan analysis , the expression level of hng fusion protein is above 40 % and m - insulin fusion protein above 50 % . western - blot result demonstrated m - insulin fusion protein had specific reaction with mouse anti human insulin antibody , we got hng fusion protein and m - insulin fusion protein with purity of above 80 %
今士考个二目卜乙成功构建了ptxbi一hng及ptxbi一m一insulin原核表达克隆，并获得了高效表达，经过纯化得到纯度人于80 %的融合蛋白，并对人胰岛素突变体融合蛋白进行了初步活性测定。 |
| 7. | Objective : construct high - level expression system of echistatin in e . coli methods : obtain amino - acid sequence of echistatin from genebank database . considering the bias of usage of 61 available aminoacid codons in e . coli , design the coding sequence of echistatin , synthesize the dna sequence chemically , get single copy coding gene and repeated two copy coding gene of echistatin . insert the sequence into expression vector pbv220 , and more , we construct fusion expression clone of echistatin with pcr , identify the recombinant vector by dna sequencing
目的构建蛇毒锯鳞蝰素( echistatin )的原核高效表达体系方法由genebank数据库检索蛇毒锯鳞蝰素( echistatin )的氨基酸序列，结合大肠杆菌蛋白质合成体系对氨基酸密码子使用的偏爱性，设计了echistatin编码基因，体外人工合成编码基因dna片段，通过适当的限制性内切酶位点插入表达载体pbv220 ，分别构建了echistatin的单拷贝表达克隆、双拷贝串联表达克隆；进一步通过pcr技术构建echistatin的融合表达基因克隆。 |
| 8. | Purpose 1 construction of prokaryotic high expression vector of human platelet factor 4 ( h pf4 ) 2 expression and purification of r h pf4 3 bioassay of r h pf4 methods according to the modulation character of eukaryotic protein expression in prokaryotic cells , we design a pair of particular primers , and construct a prokaryotic expression vector pbv220 - r hpf4 by dna polymerase chain reaction ( pcr ) and dna recombinant technic . the expression plasmid was identified with pcr and dna sequencing . pbv220 - r hpf4 was transformed into e . coli dh5a , bl21 ( de3 ) and induced by increasing the temperature to 42 . we identified the expression protein by sds - page and western - blotting
目的1人血小板因子4 ( hpf4 )原核高效表达克隆的构建2重组hpf4的表达及分离、纯化工艺研究3重组hpf4的特性研究方法根据原核细胞表达真核蛋白的基因表达调控特点，设计合成一对特异引物，在pt7 - 7 - rpf4表达质粒的基础上，应用聚合酶链式反应( pcr )对其cdna进行改造，通过dna重组技术构建成重组hpf4原核表达质粒pbv220 - rhpf4 ，用快速pcr检测法、 dna测序分析，鉴定重组hpf4表达质粒的正确性。 |
| 9. | Western - blotting result demostrated rhpf4 had specific reaction with rabbit anti - hpf4 antibody . our system improve the expression level of r hpf4 by 80 fold compared with pt7 - 7 - r hpf4 . after purified and renatured , r hpf4 prepared by our methods has bioactivity like wide hpf4 . our study establish a stable base for further reseach of the h pf4 and provide a theoretics gist for modulative mechanism of eukaryotic protein expression in prokaryotic cells
我们构建的rhpn原核高效表达系统经m page及凝胶密度扫描分析结果表明， rhpf4表达量占菌体总蛋白量的25 30 ，较原表达克隆pt7 7 rhpf4提高了近80倍，经快速高效的包涵体分高纯化工艺和复性工艺， rhpf4具有野生蛋白活性。 |
