Volume 18, Issue 4 (Winter 2021)
Sci J Iran Blood Transfus Organ 2021, 18(4): 269-278 |
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Taghikhani F, Mohammadipour M, Mousavi Hosseini K. Expression of recombinant Stem Cell Factor fused with Leucine Zipper. Sci J Iran Blood Transfus Organ 2021; 18 (4) :269-278
URL: http://bloodjournal.ir/article-1-1333-en.html
URL: http://bloodjournal.ir/article-1-1333-en.html
Abstract: (1300 Views)
Abstract
Background and Objectives
Stem cell factor (SCF) is a blood cytokine that plays an important role in the differentiation of hematopoietic progenitor cells, as well as survival, proliferation, and differentiation of mast cells. Furthermore, it can enhance tumor cell proliferation and invasion. Recombinant or natural form of leucine zipper can cause homo- or heterodimerization of many types of proteins which are attached to it and have a favorable effect on their function or stability. The present study investigates the expression of leucine zipper-bound SCF.
Materials and Methods
This experimental study consists of designing a recombinant protein by bioinformatic softwares and the laboratory part of the study. The indices of interest for designing of the protein structures were analysed by various softwares including PSIpred and Chimera and the recombinant protein stability by RAMPAGE. The sequence synthesis was performed after determination of the enzymatic cleavage sites with NEBcutter. Then, codon optimization for Origami was done with Optimizer. Cloning in TOP10 and the protein expression in Origami were performed.
Results
The results of bioinformatics studies on the Leucine Zipper/SCF recombinant protein included the necessary criteria for the possible native structure of the protein and the high score for viability in the living cell. Furthermore, the results of the experimental section showed successful cloning and expression.
Conclusions
In the present study, the results indicated the successful cloning and expression of the recombinant Leucine Zipper/SCF protein.
Background and Objectives
Stem cell factor (SCF) is a blood cytokine that plays an important role in the differentiation of hematopoietic progenitor cells, as well as survival, proliferation, and differentiation of mast cells. Furthermore, it can enhance tumor cell proliferation and invasion. Recombinant or natural form of leucine zipper can cause homo- or heterodimerization of many types of proteins which are attached to it and have a favorable effect on their function or stability. The present study investigates the expression of leucine zipper-bound SCF.
Materials and Methods
This experimental study consists of designing a recombinant protein by bioinformatic softwares and the laboratory part of the study. The indices of interest for designing of the protein structures were analysed by various softwares including PSIpred and Chimera and the recombinant protein stability by RAMPAGE. The sequence synthesis was performed after determination of the enzymatic cleavage sites with NEBcutter. Then, codon optimization for Origami was done with Optimizer. Cloning in TOP10 and the protein expression in Origami were performed.
Results
The results of bioinformatics studies on the Leucine Zipper/SCF recombinant protein included the necessary criteria for the possible native structure of the protein and the high score for viability in the living cell. Furthermore, the results of the experimental section showed successful cloning and expression.
Conclusions
In the present study, the results indicated the successful cloning and expression of the recombinant Leucine Zipper/SCF protein.
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