重组克隆

目的：建立PCR扩增快速筛选重组克隆的方法。

Aim : To set up a new method for rapid screening of recombinant clone .

小鼠Endostatin基因重组克隆的构建及鉴定

Construction of Mouse Endostatin Gene Recombinant Clone and its Identification

一种快速提取质粒DNA用于鉴别重组克隆的方法

A Rapid Method for Preparation of Plasmid DNA for Screening Recombinant Clones

单菌落PCR法直接快速鉴定重组克隆

Rapid Characterization of Recombination Clone by PCR Screening of Individual Bacterial Colonies

间日疟原虫DNA重组克隆的酶切鉴定和DNA转印杂交分析

Identification and analysis on Plasmodium vivax DNA fragments cloned by restriction endonuclease digestion and Southern blot hybridization

结论：菌落PCR可用于筛选阳性重组克隆。

Conclusion : Bacteria colony-PCR can be used in screening positive recombinant colonies .

基因变构IL-2重组克隆的构建

Construction of recombinant clone of site-specific gene mutant of human interleukin-2

将每种型别的MY09/11引物扩增区的PCR产物进行纯化、重组克隆。

PCR products of each type of MY09 / 11 primer amplification area were purified and recombinant cloned .

重组克隆的鉴定证明这个大分子cDNA确是病毒RNA3′区的拷贝。这将有利于对这类RNA病毒基因组的结构-功能分析和对病毒cDNA拷贝的感染性的研究。

Ft is advantageous for analysis of structure-function of large RNA virus and research of infection of virus cDNA copy .

通过克隆缩减产物的PCR扩增产物，得到一个包含350个重组克隆的SSH文库。

The PCR products amplified using the template from the subtractive hybridization were cloned and a SSH library containing 350 recombinants was constructed .

从入门文库中挑取1×107重组克隆，利用经LR重组反应构建cDNA表达文库。

Then a cDNA expression library was constructed through LR recombination reaction with 1 × 107 cfu picked from the entry library .

主要研究结果如下：1.应用SSH技术成功构建了小麦条锈菌诱导的cDNA文库，获得6972个重组克隆。

Main results could be summarized as follows : 1 . Rust pathogen-induced SSH cDNA library was successfully constructed and 6972 recombinant clones were preserved .

重组克隆PsecTaq2AAMG酶谱分析与预期结果一致，序列测定结果与GenBank中的人釉原蛋白序列完全一致。

The recombinant plasmid PsecTaq2A_AMG was analyzed by restriction endonuclease mapping and DNA sequencing . The results of sequencing were consistent with those from GenBank .

用X-gal平板及质粒图谱分析方法筛选重组克隆株，再用限制性核酸内切酶酶切图谱分析鉴定。

The recombinant clone was screened by X-gal plate and plasmid map , and identified by restricted enzyme mapping analysis method .

经过SDS-PAGE蛋白电泳、CO结合差光谱分析结果证明，重组克隆表达的羟化酶分子量约45kD、重组的蛋白量约占细胞总蛋白的12%，并具有细胞色素P-450的特征。

The results by SDS-PAGE and carbon monoxide-binding difference spectra analysis showed that the clone expressed C-14 hydroxylase whose molecular weight was about 45 kD , accounted for a 12 % of total proteins in the cells and possessed the cytochrome P-450 character .

结果获得了编码合信号顺序的MASP－2肽链全长cDNA，将其与pGEM－T载体连接，转化大肠杆菌TG1，建立了MASP－2的重组克隆。

Results The cDNA fragment encoding MASP-2 polypeptide with the signal sequence was isolated , linked with pGEM-T vector and transformed into E. coli TGI .

方法应用重组克隆表达抗原的血清学技术（SEREX），用异体HCC血清和非HCC血清检测两种新的HCC抗原的特异性。

Methods Serological identification of antigens by recombinant expression cloning technique ( SEREX ) was used in order to identify specificity of two HCC antigens in allogenic HCC sera and other sera including gastric carcinoma , breast cancer and normal individuals .

【结论】本实验获得Sj26GST重组克隆，为进一步研制基因工程蛋白疫苗作准备。

Conclusions GST recombinant clone was achieved in the experiment . It is prepared for further study on Gene-engineering vaccine .

方法以牵引丝蛋白基因单体为基础，利用同尾酶聚合法构建含牵引丝蛋白基因单体六聚物的重组克隆质粒pUC18-6S和重组原核表达质粒pET-28a（+）-6S。

Methods Based on the monomer of the gene of spider dragline silk protein , the recombinant plasmids pUC-6S and pET-28a ( + ) - 6S were cloned by isocaudarner enzymatic polymerization .

结论成功构FOLR-1的重组克隆及真核表达载体，为今后利用FOLR-1进行卵巢上皮性肿瘤的免疫及导向治疗研究打下了基础。

Conclusion The recombinant human FOLR-1 gene clone has been established successfully and thus provides a basis for further research of the immunotherapy or targeting therapy for ovarian carcinoma .

利用寡核苷酸探针快速筛选重组克隆

A Rapid Method of Using Oligonucleotide Probe to Screen Recombinant Clone

方法：重组克隆，基因转染及RT－PCR。

Method s : DNA recombination , gene transfection and RT-PCR .

牛生长激素基因的人工合成、重组克隆及高效表达

Synthesis , Cloning and High-level Expression of the Bovine Growth Hormone Cenes

提出了利用寡核苷酸探针快速筛选重组克隆的方法，在数小时内即可完成杂交和自显影的过程。

A rapid method of using probe to screen recombinant clone was established in this paper .

结论：成功构建了编码卫氏并殖吸虫特异性抗原的重组克隆Pw-2。

Conclusion : The recombinant plasmid Pw 2 encoding Pw specific antigen was obtained successfully prepared .

然后以重组克隆载体质粒为模板，扩增β-防御素5基因成熟肽片段。

Then , the β - defensin 5 gene mature peptide fragment was amplified according the recombinant clone vector plasmid .

结论：该方法可以快速、准确的筛选重组克隆，并能够确定片段插入的方向和大小。

Conclusion : This method could screen recombinant clone rapidly , while identifying the orientation and length of the inserted sequence .

第3轮淘筛后洗脱下来的噬菌体，较第1轮增加了近100倍。含有抗体重链基因和轻链基因的重组克隆，也由淘筛前的25%增至100%。

The eluted phage was enriched nearly 100 fold , and the percentage of recombinant clones increased from 25 % to 100 % after three rounds of panning .

随机酶切重组克隆显示，插入片段达到预期的15~23kb之间，提示该方法适于植物材料高质量基因组文库的构建，有利于分离完整的基因序列及其调控区。

Quality detection showed that the DNA inserts were between 15 ~ 23 kb as expect , suggest that the method was appropriate for constructing plant genomic library , and was helpful to isolate full-length gene and its regulation sequences .

目的应用以菌落为模板的聚合酶链反应（PCR）技术筛选插有小鼠Doc1R基因组序列的重组阳性克隆。

Objective To screen the Doc 1R gene recombinant plasmid by use of colony PCR .