Generating Cyan Fluorescent Protein (CFP) From Green Fluorescent Protein (GFPUV) By Y66w Mutation
Journal Title: IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB - Year 2018, Vol 4, Issue 1
Abstract
This work seeks to generate CFP from GFPuv by Y66W mutation. Specific objectives include site directed mutagenesis on GFPuv and sequence alignment of generated CFP with wild type GFPuv and fluorimetry studies of CFP in relation to GFPuv. Several approaches that include plasmid digestion and GFPuv ligation, colony PCR,super-competent cell transformation, sequencing, sequence alignment, Ni-NTA chromatographic purification and fluorimetry were employed. Results show that GFPuv was successfully mutated to CFP as indicated on position 65 – 67 of aligned sequence thus, a clear confirmation that Y66W mutation in GFPuv generates CFP. Fluorimetry report 22.5nm difference in peak fluorescence between the proteins implying there was a stock shift.
Authors and Affiliations
Anakaa T. M, Emediong N. E, Gloria O. O
Keywords
Related Articles
- EP ID EP415006
- DOI 10.9790/264X-04011719.
- Views 201
- Downloads 1
Role of high sensitive C–reactive protein in asthma patients
Asthma is a clinical condition characterized by inflammation of the airways and C-reactive protein is a marker for inflammation, infection, tissue damage and host defence mechanism by complement pathway. Impaired respira...
Iron Deficiency Anemia In Pregnancy: Pre And Post Iron Sucrose Therapy In A Tertiary Referral Centre
Introduction: Iron deficiency is commonly prevalent in women of reproductive age group.Early detection of iron stores before the development of anemia is beneficial. The maternal iron level is important for the growth of...
Relevance of Potential Microorganisms for Enhancing Biogas Production
Bacteria were isolated from the sewage water for the purpose of enhancing biogas production. The sample was serially diluted up to 10-7 dilution, for the isolation of hydrolytic, acidogenic, acetogenic and methanogenic b...
Bioinformatics: Application of Information Technology in Biological Science
Bioinformatics was applied in the creation and maintenance of a database to store biological information at the beginning of the "genomic revolution", such as nucleotide and amino acid sequences. Development of this type...
Efficacy of Annona squamosa (L.) extracts in controlling Diabetes Mellitus in Mice diabetic models
Diabetes mellitus (DM) is a metabolic disorder of multiple etiologies characterized by absolute or relative deficiency of insulin secretion with or without varying degree of insulin resistance. Sedentary life style and o...