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

the pESP-2 vector results in a seamless junction of the enterokinase site and the initiation codon of the gene to be expressed. The pESP-2 vector is supplied in a form ready for LIC cloning, with 12- and 13-nucleotide, single-stranded overhangs. The gene of interest is prepared for PCR amplification and LIC cloning by using primers that include 12- and 13-nucleotide sequences that are complementary to the 5 single-stranded overhangs of the pESP-2 vector. Following PCR amplification, the PCR product is treated with Pfu DNA polymerase in the presence of dATP. In the absence of dTTP, dGTP and dCTP, the 3- to 5-exonuclease activity of Pfu DNA polymerase removes 12 to 13 nucleotides at the respective ends of the PCR product. This activity continues until the first adenine is encountered, when the polymerase activity of the enzyme becomes dominant, producing a DNA fragment with 5-extended single-stranded tails that are complementary to the single-stranded tails of the pESP-2 vector. The insert anneals to the vector without further enzymatic treatment and is ready for transformation into XL10-Gold ultracompetent cells.

Protein Expression and Purification

Figure 3A & 3B

We have expressed and purified a number of recombinant proteins in S. pombe, using the pESP-2 vector and the LIC cloning strategy to create the constructs. The recombinant proteins include human Elk1, Krs2, MEK1WT and the chicken calmodulin gene product. The yields for GST-Elk1 and GST-Krs2 fusion proteins were relatively low, approximately 1 mg or less per liter, while the GST-MEK1WT
'"/>

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:1/15/2014)... Village, Calif. (PRWEB) January 15, 2014 ... treatment and management through the genomic analysis of tumor ... offer Therapy Finder™, a web-based cancer decision support application ... reporting option with Cynvenio’s ClearID™ genomic test, Therapy Finder ...
(Date:1/15/2014)... Toronto, Canada (PRWEB) January 15, 2014 ... perspectives on nonclinical and clinical safety assessment in biosimilars. ... a path for biosimilar drug development, however the complex ... regards to quality, safety and efficacy extremely challenging. Based ...
(Date:1/15/2014)... 15, 2014 Two champions of science, ... sponsorship of an annual competition for middle and high ... to, innovative STEM study. The competition presents students with ... of Engineering Aptitude, Mathematics, and Sciences is a ...
(Date:1/15/2014)... 2013 was a banner year of continued ... saw continued independent research led by the team at ... $1 million grant from the Susanne Marcus Collins Foundation, ... peer reviewed journal, Amy Grant highlighted Brainwave Optimization® in ...
Breaking Biology Technology:Cynvenio Offers New Breast Cancer Therapy Finder Application for its ClearID Breast Cancer Genomic Test 2Cynvenio Offers New Breast Cancer Therapy Finder Application for its ClearID Breast Cancer Genomic Test 3Xtalks Life Sciences Webinar Examines Safety Assessment of Biosimilars 2Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 2Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 3Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 4Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 5Dynamic Innovative Technology Showcased at Scottsdale Company’s Open House 2Dynamic Innovative Technology Showcased at Scottsdale Company’s Open House 3
... Inc. (NTI(R)) (Nasdaq: NTII ), today announced ... GmbH (Merz) for sales,by Merz and its marketing ... disease for the quarter ended September 30,2007. Under ... payments on sales of Memantine by Merz and ...
... improved but still flawed government-wide plan for nanotechnology ... system. Federally-funded studies essential to managing possible ... by a top-down strategy tied to projected commercialization, ... decision-making process. , The National Nanotechnology ...
... collaboration between researchers at the National Institute of Standards ... Kentucky potentially offers a low-cost way to mass produce ... for a wide range of applications including flexible displays, ... field diagnostics. , In a paper posted this week,* ...
Cached Biology Technology:Neurobiological Technologies Reports $2.1 Million Quarterly Payment for Sales of Memantine 2Federal nanotech risk research plan still comes up short 2Directed self-ordering of organic molecules for electronic devices 2
(Date:7/9/2014)... common frog is one of the amphibians with the ... in permanent areas of water where it comes into ... its larvae. Research carried out by the Spaniard Germn ... the larvae of these frogs have developed a defensive ... tails and larger bodies if they co-exist with the ...
(Date:7/9/2014)... measurement results accrue. To completely and systematically archive ... In fact, researchers in the life sciences spend ... to an online survey of 70 people working ... for Applied Information Technology FIT in Sankt Augustin. ... no centralized or structured approach to data collection ...
(Date:7/9/2014)... inhibitors and N-methyl-D-aspartate receptor antagonists can alleviate ... to affect irreversible cognitive dysfunction and effectively ... Amyloid beta peptide (Aβ) vaccines reduced and ... (AD) transgenic mouse model, and significantly improved ... and his team, First Affiliated Hospital of ...
Breaking Biology News(10 mins):Frogs have developed rapid defences against the red swamp crayfish 2Frogs have developed rapid defences against the red swamp crayfish 3Managing the data jungle 2Managing the data jungle 3
... of vertebrate species, but while groups such as mammals, birds ... decades, knowledge about relationships among many types of fishes was ... of scientists led by Richard Broughton, associate professor of biology ... of Oklahoma, published two studies that dramatically increase understanding of ...
... proportion of black carbon created during combustion will remain in ... burying black carbon in the ground in order to restrain ... a part of black carbon will dissolve from soil to ... to the ocean was estimated in a research article published ...
... the University of Birmingham, BGI and its open-access ... the UK,s Natural Environment Research Council (NERC) to ... large-scale environmental metabolomics data. Metabolomics involves the ... living organisms and can provide an indication of ...
Cached Biology News:Something's fishy in the tree of life 2'Black carbon' flowing from soil to oceans 2Collaboration to establish new computational resources for metabolomics 2
RABBIT ANTI HUMAN TRRAP (CT) Immunogen: Recombinant C-terminal kinase-related domain of human TRRAP....
RABBIT ANTI HUMAN CHYMOTRYPSIN...
MSE ANTI CAN. CD3:FITC/RAT ANTI CD4:RPE...
MOUSE ANTI POLY (ADP-RIBOSE) Immunogen: Purified poly (ADP-ribose) polymer, 10-50 unit chain length...
Biology Products: