双酶切

应用双酶切/Southern杂交方法诊断面肩肱型肌营养不良

Diagnosis of facioscapulohumeral muscular dystrophy using double enzyme digestion associated Southern blotting method

通过质粒双酶切和DNA测序证实该重组质粒构建正确。

This was confirmed by cleavage of restriction endonuclease and DNA sequencing .

双酶切法克隆肺炎支原体DNA的研究

Study on cloning of Mycoplasma pneumoniae DNA digested with double restriction enzymes

PCR、双酶切鉴定以筛选PET15b-HSP65、PET15b-HSP70阳性重组表达质粒。

The recombinant plasmids are identified by PCR and double digestion .

经限制性内切酶双酶切鉴定及DNA测序证实此蛋白共表达载体构建成功。

Enzyme digestion analysis and DNA sequencing showed that the co-expressed protein expressing vectors were constructed successfully .

分别经双酶切、特异PCR及测序法对重组质粒进行鉴定。

The recombinants were identified with double GA endonuclease digestion , specific PCR and sequencing .

并对重组质粒进行双酶切、菌落PCR及测序分析进行鉴定。

To verified the by restriction enzyme digestion , bacterial colony PCR , and sequencing analysis .

这两次转化过程，均经过菌落PCR和限制性内切酶双酶切检验。

For both cloning vectors , positive clones were verified by colony PCR and restriction enzyme double digestion .

结果：经PCR和双酶切鉴定，重组表达载体pQE30/P34H为正确克隆。

Results : Recombinant expression vector pQE-30 / P34H was correctly constructed , identified with PCR and double-enzyme digestion .

AFLP是一种新的DNA标记，它是利用PCR技术对基因组DNA双酶切片段的选择性扩增。

AFLP a newly DNA markers , involves selective amplification of a subset of restriction fragments generated by double digestion of genomic DNA .

对其分析过程中DNA提取、双酶切、连接、预扩增、标记和选择性扩增的效果进行了检测。

The results of several reactions during AFLP analysis have been detected such as DNA extraction , double enzymes restriction , adapter ligation , pre amplification , labeling and selective amplification .

将该基因片段插入T／A克隆，经PCR、双酶切和测序鉴定，获阳性克隆加以扩增，试剂盒抽提阳性克隆。

The above fragment was inserted into T / A vector and identified by PCR , enzyme digestion and gene sequencing . The positive clone identified was amplified .

结果经双酶切、PCR及测序鉴定证实血凝素基因HA1区的真核表达载体构建成功。

By restriction enzyme identification , PCR and sequence analysis , the recombinant plasmid of the HA1 was successfully constructed .

探讨和初步建立了利用双重PCR和双酶切技术同时检测ESR、RYR1两个基因的变异的方法。

The duplex PCR and double digestion technique were established to detect polymorphisms of ESR and RYR1 simultaneously .

将这2个双酶切产物连接并转化，通过酶切鉴定和PCR鉴定证明完成了vp4原核表达载体的构建。

The completion of the construction of the vp4 prokaryotic expression vector was proved through the identification of enzyme-cutting and PCR analysis .

结果PCR产物和构建的重组质粒pET22b-p24经双酶切插入的外源基因片段均为690bp，与预期p24抗原全基因片段大小一致。

Results PCR product and external gene section from the recombinant plasmid pET 22b-p24 showed the same size of 690 bp equal to p24 gene sequences .

并运用双酶切技术、SDS-PAGE电泳、westernblot（WB）及ELISA法分别对插入基因片段的正确性、表达产物的活性及特异性进行检测。

The accuracy of inserted gene and activity , specificity of HIV p24 proteins were detected by two enzymes digestion technology , SDS-PAGE , Western Blot ( WB ) and ELISA .

然后利用限制性内切酶SfiⅠ、NotⅠ分别双酶切抗体基因和噬菌体展示载体。

Then the antibody genes and the phage display vector were digested by restriction endonucleases Sfi ⅰ and Not ⅰ .

扩增产物经双酶切后，重组到表达载体pET-32a（+）中。

The PCR product was inserted into expression plasmid pET-32a ( + ) after restriction digest .

结果构建的EBOG87和EBOWT重组载体经内切酶双酶切鉴定及核苷酸序列测定证实。

Results The constructed vectors of EBO-WT and EBO-G87 were identified by restriction enzyme digestion and nucleotide sequencing .

结论PCR双酶切法检测CMT1A基因重复敏感、快速、准确，适于临床推广应用；

Conclusion The PCR combined with restriction enzyme digestion represented a relatively sensitive and accurate method for detecting gene duplication in CMT1A cases for clinical diagnosis .

在进行连锁群步移的基础上，建立了基于BAC文库、双酶切和Southern杂交的构建线粒体DNA物理图谱的体系。

In order to isolate and identify the CMS genes , we developed a new platform to construct physical map of chromosome DNA by means of restriction enzyme double digestion . The elongation of contigs is based on Southern hybridization .

PCR产物经过双酶切后按照正确的读码框顺序克隆到毕赤酵母表达载体pPICZαA中；

PCR products was inserted into the plasmid pPICZ α A after digested by the Xho ⅰ and Xba ⅰ restriction enzymes according to the correct reading code frame ;

随机挑选单菌落小规模培养，碱裂解法提取质粒DNA，PstⅠ／BglⅡ双酶切后行变性聚丙烯酰胺凝胶电泳和银染分析，筛选含有不同等位基因的重组质粒后测序。

Use alkaline lysis method to extract the plasmid DNA . DNA constructs were identified by Pst I / Bgl II digestion , denaturing polyacrylamide gel electrophoresis and DNA sequence analysis .

经PCR、双酶切及测序鉴定后将重组质粒PET-32a（+）-gI转化入大肠杆菌BL21（DE3）感受态细胞进行表达。

After being confirmed by PCR , restriction endonuclease digestion and sequencing , pET-32a ( + ) - gI was transformed into E.coli BL21 ( DE3 ) competent cells for overexpression .

双酶切及测序结果表明：目标基因已成功插入表达载体pET-32a（+）。

Restriction endonuclease digestion analysis and sequencing data revealed that the target gene has been successfully inserted the expression vector pET-32 a ( + ) .

本研究构建了大菱鲆扩增片段长度多态性（AFLP）分析体系，对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。

AFLP ( Amplified Fragment Length Polymorphism ) analysis system for turbot was established in this study , with the relative processes being presented , including DNA extraction , double enzymes digestion reaction , adapter ligation reaction , pre-amplification and selective amplification reactions , and argent dyeing .

方法：通过双酶切把PScDNA全长序列接到pBlueskriptⅡKS（+）的多克隆位点上；

Methods : Link the PS cDNA full-length sequence to the multiple cloning site of pBlueskript ⅱ KS ( + ) by double restriction enzyme digest .

将合成后的基因序列经NdeI和HindIII双酶切，并与经同样酶切的pET-22b表达载体进行连接，通过PCR扩增、双酶切鉴定及核苷酸测序证实原核表达载体构建成功。

( 2 ) The pET-22b expression vector was chosen , and Nde I and Hind III restriction sites were selected . The prokaryotic expression vector was confirmed to be constructed successfully by restriction enzyme digestion , PCR and sequencing .

经双酶切及DNA测序鉴定后，两种真核表达重组体均成功构建，分别命名为truncated-ATP、truncated-ATP-CT。

After the identification by two restriction enzymes digestion and DNA sequencing , both of two eukaryotic expression recombinants are to be successfully constructed , which named truncated-ATP and truncated-ATP-CT .