Navigation Links
New Yeast Cloning System for Producing Proteins with Native Amino Acid,,,Sequences

es of PCR) and Stratagenes high-fidelity Pfu DNA polymerase. After purification of the PCR product, a simple, quick Pfu DNA polymerase treatment of the PCR product replaces restriction enzyme digestion; annealing of insert and vector fragments with specific ends ensures high cloning efficiency. Cloning of the PCR product is directional, and more than 80% of the transformants contain an insert in the correct orientation.

The PCR products are cloned into the pESP-2 vector immediately downstream of the GST affinity tag and the enterokinase (EK) recognition site, resulting in a seamless junction between the EK cleavage site and the initiation codon (ATG) of the gene to be expressed. After expression in S. pombe, the GST fusion protein can be purified from crude cell extracts using a single-step purification procedure based on the selective affinity of the GST peptide for glutathione immobilized on resin. Following the single-step purification procedure, the GST affinity tag may be removed by proteolytic cleavage with EK to yield polypeptides with native amino acid sequence. Cleavage with thrombin yields the recombinant protein fused with the FLAG epitope for future analysis of the protein.

The ESP LIC cloning kit contains pretreated, ready-to-use pESP-2 vector, Pfu DNA polymerase and 10X reaction buffer, Epicurian Coli XL10-Gold ultracompetent cells, forward and reverse primers for sequencing and PCR amplification and other LIC cloning reagents and control plasmids. The ESP LIC cloning and protein purification kit includes all of these components plus S. pombe competent cells (strain SP-Q01), YES media for vegetative growth, Edinburgh minimal media (EMM) for induction, acid-washed glass beads and GST affinity resin.

Ligation-Independent Cloning

figure 2

The LIC strategy (figure 2) using
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Epitope-Tagging Vectors for Functional Analysis in Yeast
2. Yeast Protein Production System Features High Yields and One-Step Purification
3. Electroporation of Yeast Artificial Chromosomes
4. New Automated Yeast Cell Counter Ends Tedium & Errors of Manual Counting
5. High Yield Protein Production from Pichia pastoris Yeast: A Protocol for Benchtop Fermentation
6. Enhanced PCR Cloning System
7. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
8. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
9. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
10. QIAGEN PCR CloningPlus Kit
11. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
Post Your Comments:
(Date:8/21/2014)... The College of New Rochelle announced ... the Mid-Hudson Region to be designated as a START-UP ... from qualified “high-technology” businesses that align with the College’s ... have been selected for the START-UP NY program and ... stimulate economic development in New York by growing business ...
(Date:8/21/2014)... NEW YORK , Aug. 21, 2014 /PRNewswire/ ... OTCM) congratulates Immune Pharmaceuticals Inc., a biotechnology company, ... Capital Market.  Immune previously traded on OTCQX®, the ... operated by OTC Markets Group. Logo - ... Immune Pharmaceuticals on the successful execution of its ...
(Date:8/21/2014)... Pursuit Solutions ( http://www.pursuit-solutions.com ), ... services for Life Sciences sales and marketing organizations, ... New Commercial Sales Models” by Peter Robinson, co-founder ... article examines some innovative models used to improve ... in the pharmaceutical market. Robinson grades the industry ...
(Date:8/21/2014)... MA (PRWEB) August 21, 2014 ... company, has been recognized as one of the ... | 5000 magazine for the 2nd consecutive year. ... its 33rd annual Inc. 500|5000, an exclusive ranking ... represents the most comprehensive look at the most ...
Breaking Biology Technology:The College of New Rochelle Becomes First Private College in Mid-Hudson Region to Earn START-UP NY Designation 2The College of New Rochelle Becomes First Private College in Mid-Hudson Region to Earn START-UP NY Designation 3OTC Markets Group Congratulates Immune Pharmaceuticals on NASDAQ Listing 2Pursuit's Peter Robinson Publishes "Grading Pharma's Use of New Commercial Sales Models" 2SoundConnect Returns on Inc. 5000 List of Fastest Growing Companies 2SoundConnect Returns on Inc. 5000 List of Fastest Growing Companies 3
... , ... , , , ... in the homeostasis of cell numbers in adult,organisms, furthermore in eliminating ... two distinct ways of,eukaryotic cell death: oncosis and apoptosis. How cells,die ...
... , , ... , , , ... and thymidylate,synthase (TS) play a pivotal role in the metabolism,of ... is the standard,chemotherapy treatment for a wide range of solid,tumors. ...
... , , ... , Since the first description of ... a few years ago, RNAi has become a powerful,tool for analyzing ... that are,homologous to the suppressed gene. These small interfering,RNAs (siRNAs) of ...
Cached Biology Technology:Quantification of Nucleosomes in Serum,by the Cell Death Detection ELISAplus 2Quantification of Nucleosomes in Serum,by the Cell Death Detection ELISAplus 3Quantification of Nucleosomes in Serum,by the Cell Death Detection ELISAplus 4Quantification of Nucleosomes in Serum,by the Cell Death Detection ELISAplus 5Quantification of Nucleosomes in Serum,by the Cell Death Detection ELISAplus 6Gene-Expression Analysis of TP, DPD, and TS Using,the LightCycler mRNA Quantification KitsPLUS 2Gene-Expression Analysis of TP, DPD, and TS Using,the LightCycler mRNA Quantification KitsPLUS 3Gene-Expression Analysis of TP, DPD, and TS Using,the LightCycler mRNA Quantification KitsPLUS 4Quantification of siRNA Silencing Efficiency,Using the LightCycler System 2Quantification of siRNA Silencing Efficiency,Using the LightCycler System 3Quantification of siRNA Silencing Efficiency,Using the LightCycler System 4Quantification of siRNA Silencing Efficiency,Using the LightCycler System 5
(Date:8/20/2014)... a study of 158 pregnant teenagers in Rochester, NY, ... intentional consumption of ice, cornstarch, vacuum dust, baby powder ... Cornell study. , Moreover, such teens had significantly lower ... eat nonfood substances. , Pregnant teens, regardless of pica, ... lead to iron deficiency and anemia. Low iron in ...
(Date:8/20/2014)... life can persist in a cold, dark world. A ... that examined waters and sediments from a shallow lake ... extreme environment supports microbial ecosystems. , The National ... has implications for life in other extreme environments, both ... are published in the current edition of the science ...
(Date:8/20/2014)... Arizona State University, the Wellcome Trust Sanger Institute and ... isolated Mycobacterium pinnipedii from skeletons found in ... pathogen is a relative of the TB bacterium that ... today. These researchers assume that seals carried the pathogens ... sea lions was unexpected" comments Sebastien Gagneux, from the ...
Breaking Biology News(10 mins):Pica in pregnant teens linked to low iron 2University of Tennessee research uncovers subglacial life beneath Antarctic ice sheet 2
... disease affecting a person,s ability to coordinate and control ... in a finger will eventually lead to difficulty in ... without assistance. Since the 1950s research has shown that ... dopamine in their brains, which is involved in sending ...
... in rehabilitating goats and other animals that have suffered from ... In this first scientific study of rescued animals, ... had endured poor welfare, such as inappropriate diet, and lack ... created a spatial awareness test, which involved giving the animals ...
... April 22, 2013 The U.S. Farmers  & Ranchers ... multiple disciplines, including farmers, media, industry and academia,  to ... discussion was moderated by CNBC Senior Analyst and Financial ... Chicago , which took place during the ...
Cached Biology News:New research examines connection between inflammatory stimulus and Parkinson's disease 2New research examines connection between inflammatory stimulus and Parkinson's disease 3Rescue me: New study finds animals do recover from neglect 2CNBC Senior Analyst Ron Insana Moderates Discussion On Biotech Seeds At The Food Dialogues(SM): Chicago 2CNBC Senior Analyst Ron Insana Moderates Discussion On Biotech Seeds At The Food Dialogues(SM): Chicago 3
RECOMBINANT EQUINE TNF ALPHA Product Type: Recombinant Protein...
Request Info...
Request Info...
1+ mg expected yield of purified protein, expressed in insect cells - excludes cDNA...
Biology Products: