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

fusion protein gave approximately 4 mg per liter (data not shown). The GST-chicken calmodulin recombinant protein produced approximately 12 mg per liter of fusion protein. Figure 3 shows the one-column purification of the GST-calmodulin fusion protein and subsequent digestion with enterokinase.

Conclusions

The pESP LIC cloning and protein purification kits allow efficient and directional cloning of high-fidelity PCR products immediately downstream of the GST purification tag and the recognition site for enterokinase. Single-step purification of the recombinant GST fusion protein and removal of the purification tag by enterokinase cleavage yield polypeptides with completely native amino acid sequences. The ESP LIC cloning and protein purification kit includes all the necessary components for LIC cloning, expression in S.pombe and GST affinity purification.

Materials and Methods

For protein induction, the expression strain was inoculated into 10 ml of YES media and incubated at 30C overnight. This culture was used to inoculate another 10 ml of YES media and was incubated at 30C until the culture reached mid-log phase. The cells were collected by centrifugation, washed once with 50 ml of sterilized water, resuspended in 10 ml of EMM media with 25 M thiamine (repressed) or without thiamine (induced) and incubated at 30C for 18 to 20 hours. Crude lysates were prepared by washing cells in PBST buffer (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4 and 1% Triton X-100), then lysing cells by vortexing with glass beads in PBST buffer containing protease inhibitors (1 mM PMSF, 1 g/ml aprotinin, 1 M pepstain A, 100 M leupeptin and 1 g/ml chymostatin). Soluble protein was bound to glutathi
'"/>

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:8/26/2015)... , Aug. 26, 2015 /PRNewswire/ - iCo Therapeutics ... today reported financial results for the quarter ended ... expressed in Canadian dollars and presented under International ... pre-clinical activities with respect to our Oral Amp ... a contract manufacturer," said Andrew Rae , ...
(Date:8/26/2015)... , Aug. 26, 2015  The Diabetes Research ... University of Miami Miller School of Medicine, announced ... trial to test for the first time a ... completed.  This FDA approved Phase I/II study builds ... and is an important first step toward the ...
(Date:8/26/2015)... BEIJING , August 26, 2015 ... DiaCardio wins 1 st place  Wayerz ... led in Israel by venture capital fund ... countries including the US, China , ... Latin America .  Impressive achievement for the Israeli ...
(Date:8/26/2015)... , August 26, 2015 ... biggest factors fueling the optimism in biotech,s forecast ... collaborations and life sciences advancements.  Active biotechnology & ... Pressure BioSciences, Inc. (OTCQB: PBIO), Sequenom, Inc. (NASDAQ: ... SRPT ), Raptor Pharmaceutical Corp. (NASDAQ: RPTP ...
Breaking Biology Technology:iCo Therapeutics Announces Second Quarter 2015 Financial Results 2iCo Therapeutics Announces Second Quarter 2015 Financial Results 3Diabetes Research Institute Successfully Transplants First Patient In Pilot BioHub Trial 2Diabetes Research Institute Successfully Transplants First Patient In Pilot BioHub Trial 3Diabetes Research Institute Successfully Transplants First Patient In Pilot BioHub Trial 4Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 2Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 3Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 4Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 5Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 6New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 2New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 3New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 4New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 5New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 6New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 7
... N.J., June 21, 2011 Ikaria, Inc., a critical ... designed to address the significant needs of critically ill ... that its Chairman and CEO, Daniel Tasse, will present ... and International Convention on Tuesday, June 28, 2011 from ...
... 21, 2011 ,   ... announce that its,US-subsidiary NextGen Sciences Inc, a leader ... first cerebrospinal,fluid (CSF) multiple protein (multiplex) assay, ... and qualification . The assay simultaneously measures,43 human ...
... successful product commercialization, the relationship between marketing and ... the launch process. In this Regulatory Affairs Professionals ... Marketing need to collaborate on product design to ... both optimal market adoption and regulatory compliance. Led ...
Cached Biology Technology:Ikaria® To Present at 2011 BIO Business Forum and International Convention 2NextGen Sciences Launches Multiple Protein cerebrospinal fluid (CSF) CNS Disease Biomarker Assay 2NextGen Sciences Launches Multiple Protein cerebrospinal fluid (CSF) CNS Disease Biomarker Assay 3How-To RAPS Webinar June 30: Regulatory and Marketing Symbiosis from Concept to Launch 2
(Date:8/5/2015)... 2015 The biosensors market is proving highly ... into newer sectors, and development of devices resulting in ... has seen the entry of multiple participants each year ... (Photo - http://photos.prnewswire.com/prnh/20150805/255570 ) ... the Global Biosensors Market ( http://www.frost.com/nee9 ), finds ...
(Date:7/31/2015)... -- Den 10. internationale konference om Genomik (ICG-10, ... 25. oktober i Shenzhen , Kina. ... år. Siden starten i 2006 er ICB blevet en ... er en af de mest dynamiske, entusiastiske og sjove ... gennembrud og fremskridt inden for ,omik,-forskning, som yderligere vil ...
(Date:7/31/2015)... julio de 2015 La 10 th International ... medio de BGI del 22 al 25 de octubre de 2015 ... Este año, la conferencia celebra su décimo aniversario. Desde su ... una de las reuniones anuales más influyentes del mundo en ... más dinámicas, entusiastas y mejores a nivel científico. ...
Breaking Biology News(10 mins):Innovative Biosensors Incite Use in Non-Traditional Applications 2Innovative Biosensors Incite Use in Non-Traditional Applications 3Innovative Biosensors Incite Use in Non-Traditional Applications 4Den 10. internationale konference om Genomics (ICG-10) åbner i oktober 2La 10th International Conference on Genomics (ICG-10) se llevará a cabo en octubre 2La 10th International Conference on Genomics (ICG-10) se llevará a cabo en octubre 3
... food meal is never good for you, but washing that ... a new University of Guelph study. Researcher Marie-Soleil Beaudoin ... levels spike after eating a high-fat meal, but that the ... coffee jumping to levels similar to those of people ...
... leaves influence the sunlight reaching inner canopy leaves by changing ... physiology of leaves within the tree canopy is not constant, ... crown. This phenomenon is expected to have important consequences for ... A new study describes how the leaves in the outer ...
... ARBOR, Mich.Getting older doesn,t mean giving up muscle strength. ... loss that comes with age, but the Golden Years can be ... Michigan Health System. "Resistance exercise is a great ... people can function more readily in daily life," says Mark Peterson, ...
Cached Biology News:Got a craving for fast food? Skip the coffee, study says 2Older and stronger: Progressive resistance training can build muscle, increase strength as we age 2Older and stronger: Progressive resistance training can build muscle, increase strength as we age 3Older and stronger: Progressive resistance training can build muscle, increase strength as we age 4
Mouse Endoglin/CD105 Fluorescein MAb (Clone 209701)...
Catalase (Control)...
Human RELT/TNFRSF19L MAb (Clone 238104)...
... assay kit measures NOS activity by monitoring ... This assay is simple, sensitive, and specific ... with both crude and purified enzyme preparations. ... for 50 total reactions. Radiolabeled arginine and ...
Biology Products: