| 1. | Application of quantum dots labeling to the interaction between reconstructed - phage display peptide and insulin receptor
量子点荧光标记在重组噬菌体表面展示肽与胰岛素受体相互作用中的应用 |
| 2. | The phagemid particles displaying functional scfvs were rescued by reinfection of helper phage m13ko7 , thus a murine antibody library was obtained
经辅助噬菌体m13ko7超感染回收全部重组噬菌体，此即噬菌体抗体库。 |
| 3. | The patterns of sds - page indicated more than 30 % of recombinant proteins could be obtained from the extract of e . coli bl21 . furthermore , library of recombinant phage antibodies was constructed from total rna isolated from spleen of the female balb / c mice immunized by bull sperm
首先用牛精子免疫雌性四周龄balb c小鼠，从其脾脏组织分离总rna ，应用重组噬菌体抗体库技术，构建了一个针对牛精子的噬菌体抗体文库。 |
| 4. | The library was rescued with phage m13k07 in order to display scfv on the surface of the phage and to form the recombinant phage antibody library . one of positive scfv clones , named pcsal , was selected with phage - elisa after panning and screening by bull sperm three times . scfv fragment , amplified from pcsa1 , was ligated to pmd18 - t vector for sequencing analysis
取阳性重组噬菌体抗体克隆株pcsa1 ， pcr扩增其scfv基因，筛选重组子进行序列测定，发现其序列符合小鼠抗体基因的一般特征，并且与几株抗磷酸胆碱的抗体重链和轻链可变区序列的同源性达80以上；推测pcsa1scfv针对的抗原是磷酸胆碱类物质。 |
| 5. | The library consisted of 1 . 3 x 106 clones with an average insert size of about 18kb . the capacity of this library was about 20 times the equivalent of the genome of atriplex centralasiatica iljin . screening the genomic library with a 400bp probe located at the 5 ' end of the badh gene obtained by rt - pcr , we got four positive clones
3x10 ‘个重组噬菌体，插入片段大小约为18kb ，含插入片段的频率为100队以中亚滨蓉甜菜碱醛脱氢酶门adh ）基因近5 ’端的约400hp片段为探针，筛选中亚滨蓉基因组文库，得到了4个阳性克隆。 |
| 6. | Detection of antigen - binding affinity of mg7 recombinant phage antibody elisa was repeated to confirm the antigen - binding affinity of positive clones screened out in the former procedure ; these positive phages were examined by restriction analysis ( ecor i and hind iii ) ; competitive elisa was performed to test the inhibitory ratio of these positive clones to the binding affinity of mg7mab and its relevant antigen , the positive dones possessing apparent inhibitory effect were singled out for later use
X阳性克到驶知烘扳原kbbjg ）浊对阳性克隆进行限制陇卧赐析（ uwi和hedlll ）鉴定三用竞争elisatoljmg7重组噬菌体抗体性克隆对mg7单扶与其相应抗原结合忙的喇率，从中j ） ed出对mg7单抗与期眩抗原结合有抑余j效应的克隆用于进一涉研究。 |
| 7. | Panning and enrichment ofmg7 recombinant phage antibody the transformed cells were infected with m13k07 helper phage to produce a phage form of mgy recombinant phage antibody library ; after three rounds panning of the library with the mgrbinding antigen highly expressed gastric cancer cell line kato iii , the mg7 scfv displayed phage dones were selected from the enriched recombinant phages by elisa
3 mg _ 7重组噬菌体抗体库的淘筛用m13ko7辅助噬菌体感染转化菌，以挽救出噬菌体形式的抗体库；用高表达mg _ 7抗原的胃癌细胞系kato对抗体库进行三轮淘筛；用elisa筛检呈现有mg _ 7scfv的噬菌体单克隆。 |
| 8. | In the present study , the express library of monoclonal anti - sp18 scfv ( single chain fragment variable ) gene is constructed and selected for further study of sp18 antigen on mammalian fertilization and embryogenesis . total rna were firstly isolated from these growing hybridoma cells which secretes monoclonal anti - sp18 antibodies . after obtained using rpas system , vh and vl genes were used to assemble scfv gene fragment with a linker primer
应用重组噬菌体抗体库技术，从分泌小鼠抗牛精子sp18抗体的杂交瘤细胞系中分离总rna ，克隆抗体重链和轻链可变区基因，加入连接肽引物( linkerprimer )组装成单链抗体scfv ( singlechainfragmentvariable )基因并用rs引物进行扩增， sfi 、 not酶切，回收后与pcantab5e载体相连，转化e . colitg1宿主菌，构建单链抗体文库。 |
| 9. | Culture of mg7 hybridoma cells and detection of antigen - binding affinity of mg7 mab by elisa 2 . construction and identification of mg7 recombinant phage antibody library mrna was isolated from cultured mg7 hybridoma cells and converted into cdna ; the variable fragments of heavy and light chain were separately amplified and assembled into scfvs with a specially constructed dna linker by pcr . the scfvs dma was ligated into the phagmid vector pcantabse and the ligated sample was transfered into competent e . co / / tg1 to generate a bacterial form of mg7recombinant phage antibody library ; volume and recombinant ratio of the library were evaluated by means of bacterial colony counts and restriction analysis ( ecor i and hind iii )
Mg _ 7重组噬菌体抗体库的构建及鉴定从培养的mg _ 7杂交瘤细胞中提取并分离mrna ，反转录成cdna ；利用pcr分别扩增mg _ 7单抗的重链及轻链可变区基因，并通过? dna连接子将二者连接起来形成mg _ 7单链抗体基因；将mg _ 7单链抗体基因插入pcantab5e ；将连接产物转化感受态tg1大肠杆菌，制备细菌形式的mg _ 7重组噬菌体抗体库；通过菌落计数和限制性酶切分析( ecor和hind )评估mg _ 7重组噬菌体抗体库的容量和重组率。 |
