نتایج جستجو برای: splicing by overlap extension pcr

تعداد نتایج: 7193854  

پایان نامه :وزارت علوم، تحقیقات و فناوری - پژوهشگاه ملی مهندسی ژنتیک وزیست فناوری 1389

ژن درمانی با استفاده از روش هدف گیری ژنی (gene targeting) یکی از بهترین روشهای درمان نقائص ژنتیکی نظیر بتا تالاسمی می باشد. بیماری بتا تالاسمی در اثر کاهش یا فقدان سنتز زنجیره بتا گلوبین به وجود می آید. یکی از روشهای درمان موثر برای این بیماری ژن درمانی توسط حامل های لنتی ویروسی می باشد. ظرفیت حامل های لنتی ویروس حدود 10 کیلو باز است. هدف از این تحقیق طراحی و ساخت حامل لنتی ویروس های نوترکیب فا...

Journal: :Antimicrobial agents and chemotherapy 2008
Koleen J Herlihy Joanne P Graham Robert Kumpf Amy K Patick Rohit Duggal Stephanie T Shi

To address the need for broad-spectrum antiviral activity characterization of hepatitis C virus (HCV) polymerase inhibitors, we created a panel of intergenotypic chimeric replicons containing nonstructural (NS) protein NS5B sequences from genotype 2b (GT2b), GT3a, GT4a, GT5a, and GT6a HCV isolates. Viral RNA extracted from non-GT1 HCV patient plasma was subjected to reverse transcription. The N...

Journal: :Genetics and molecular research : GMR 2015
J M Xi H Pang X L Hu Z J Wang

The objective of this study was to identify and construct a human natural phage single-chain antibody (scFv) library of human anaplastic thyroid carcinoma (ATC) using phage display technology. Total RNA was extracted from lymphatic tissue near an ATC and used to amplify variable heavy chain (VH) and variable light chain (VL) fragments with added linker sequences using reverse transcription-poly...

2017
Chang-Jun Liu Hui Jiang Lei Wu Ling-Yun Zhu Er Meng Dong-Yi Zhang

Here, an efficient cloning strategy for large DNA fragments and for simultaneous assembly of multiple DNA fragments assembly is presented. This strategy is named OEPR (based on Overlap Extension PCR and Recombination in vivo). OEPR cloning is a seamless, restriction- and ligation-independent method. The method takes advantage of both homologous recombination enzymes in E. coli and overlap PCR. ...

Journal: :Proceedings of the National Academy of Sciences of the United States of America 1992
J Tavernier T Tuypens G Plaetinck A Verhee W Fiers R Devos

By use of a 3' extension PCR strategy, cDNA clones were isolated spanning the transmembrane region and a complete cytoplasmic domain of the human interleukin 5 receptor alpha subunit (hIL5R alpha). These cDNAs differ from previously isolated clones encoding a soluble hIL5R alpha form by a sequence switch at position 1243. When expressed in COS-1 cells, only low-affinity binding of 125I-labeled ...

2013
Wataru Fujii Kiyoshi Kano Koji Sugiura Kunihiko Naito

Zinc finger nuclease (ZFN) is a useful tool for endogenous site-directed genome modification. The development of an easier, less expensive and repeatedly usable construction method for various sequences of ZFNs should contribute to the further widespread use of this technology. Here, we establish a novel construction method for ZFNs. Zinc finger (ZF) fragments were synthesized by PCR using shor...

2011
Yihong Cui Guangqin Zhu Qiuju Chen Yunfei Wang Mingming Yang Yuxuan Song Jiangang Wang Binyun Cao

Complementary DNA (cDNA) is valuable for investigating protein structure and function in the research of life science, but it is difficult to obtain by traditional reverse transcription. In this study, we employed a novel strategy to clone the human leukemia inhibitory factor (hLIF) gene cDNA from genomic DNA directly isolated from the mucous membrane of mouth. The hLIF sequence can be acquired...

Journal: :Molecular and cellular biology 2008
Julian P Venables Chu-Shin Koh Ulrike Froehlich Elvy Lapointe Sonia Couture Lyna Inkel Anne Bramard Eric R Paquet Valérie Watier Mathieu Durand Jean-François Lucier Julien Gervais-Bird Karine Tremblay Panagiotis Prinos Roscoe Klinck Sherif Abou Elela Benoit Chabot

Alternative splicing is a key mechanism regulating gene expression, and it is often used to produce antagonistic activities particularly in apoptotic genes. Heterogeneous nuclear ribonucleoparticle (hnRNP) proteins form a family of RNA-binding proteins that coat nascent pre-mRNAs. Many but not all major hnRNP proteins have been shown to participate in splicing control. The range and specificity...

Background: The immune antigen of Bacillus anthracis is a protein that can attach to the surface receptor of all human cells. At the surface of cancer cells, there is a receptor that activates the uPA (Urokinase plasminogen) that do not exist in normal human cells. Objectives: The aim of this study was changing the location of the attachment of the PA gene by a dir...

Background: The immune antigen of Bacillus anthracis is a protein that can attach to the surface receptor of all human cells. At the surface of cancer cells, there is a receptor that activates the uPA (Urokinase plasminogen) that do not exist in normal human cells. Objectives: The aim of this study was changing the location of the attachment of the PA gene by a dir...

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