CYTOSOLIC LOCALIZATION OF MOUSE PEROXISOMAL PROTEIN/ΔSKI FUSED WITH ENHANCED GREEN FLUORESCENT PROTEIN INTO CHINESE HAMSTER OVARY-K1 AND P19 CELLS

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OSTAD SHARIF M.; GHAEDI K.; NASRESFAHANI MOHAMMAD HOSSEIN; TANHAEI S.; KARBALAEI KH.; PARIVAR KAZEM; BAHARVAND HOSSEIN

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Journal: Cell Journal (Yakhteh) Year:2009 | Volume:11 | Issue:2 (42) Page(s): 154-159

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Title

CYTOSOLIC LOCALIZATION OF MOUSE PEROXISOMAL PROTEIN/ΔSKI FUSED WITH ENHANCED GREEN FLUORESCENT PROTEIN INTO CHINESE HAMSTER OVARY-K1 AND P19 CELLS

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Keywords

PEROXISOMAL PROTEINQ1

PEROXISOME TARGETINGQ1

SIGNAL PEPTIDEQ1

Abstract

Objective: Amino acid alignment analysis of deduced amino acid residues revealed a tripeptide (SKI) at the carboxy terminus of PEROXISOMAL PROTEIN cDNA. In order to see the importance of the above sorting signal, we have performed a site-directed mutagenesis to delete SKI tripeptide and its transfection into CHO-K1 and P19 cells.Materials and Methods: In order to create the appropriate site-directed mutant, PCR was done with specific primers. Amplified PeP cDNAs either containing SKI or deleted ones were constructed downstream of EGFP cDNA under regulation of the cytomegalovirus (CMV) promoter in a pEGFP-C1 vector. Transfection of P19 and CHO cells was done with lipofectamine2000.Results: Gradient PCR showed that the best annealing temperature was 71.6oC. Transfection of plasmids containing chimera of EGFP-PEP cDNAs into the CHO-K1 and P19 cells showed several punctuate structures, presumably peroxisomes, while SKI deletion showed a cytosolic mislocalization of the EGFP pattern.Conclusion: Taken together, these data strongly suggest that SKI, which is located at the C- terminus of the protein, is required for sorting this protein.

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APA: Copy

OSTAD SHARIF, M., GHAEDI, K., NASRESFAHANI, MOHAMMAD HOSSEIN, TANHAEI, S., KARBALAEI, KH., PARIVAR, KAZEM, & BAHARVAND, HOSSEIN. (2009). CYTOSOLIC LOCALIZATION OF MOUSE PEROXISOMAL PROTEIN/ΔSKI FUSED WITH ENHANCED GREEN FLUORESCENT PROTEIN INTO CHINESE HAMSTER OVARY-K1 AND P19 CELLS. CELL JOURNAL (YAKHTEH), 11(2 (42)), 154-159. SID. https://sid.ir/paper/16407/en

Vancouver: Copy

OSTAD SHARIF M., GHAEDI K., NASRESFAHANI MOHAMMAD HOSSEIN, TANHAEI S., KARBALAEI KH., PARIVAR KAZEM, BAHARVAND HOSSEIN. CYTOSOLIC LOCALIZATION OF MOUSE PEROXISOMAL PROTEIN/ΔSKI FUSED WITH ENHANCED GREEN FLUORESCENT PROTEIN INTO CHINESE HAMSTER OVARY-K1 AND P19 CELLS. CELL JOURNAL (YAKHTEH)[Internet]. 2009;11(2 (42)):154-159. Available from: https://sid.ir/paper/16407/en

IEEE: Copy

M. OSTAD SHARIF, K. GHAEDI, MOHAMMAD HOSSEIN NASRESFAHANI, S. TANHAEI, KH. KARBALAEI, KAZEM PARIVAR, and HOSSEIN BAHARVAND, “CYTOSOLIC LOCALIZATION OF MOUSE PEROXISOMAL PROTEIN/ΔSKI FUSED WITH ENHANCED GREEN FLUORESCENT PROTEIN INTO CHINESE HAMSTER OVARY-K1 AND P19 CELLS,” CELL JOURNAL (YAKHTEH), vol. 11, no. 2 (42), pp. 154–159, 2009, [Online]. Available: https://sid.ir/paper/16407/en

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