CONSTRUCTION OF MOUSE PEP CDNA AND EGFP IN TET OFF EXPRESSION VECTOR

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MONTAZERI NAJAFABADI BEHSHAD; GHAEDI KAMRAN; DORMANI KIANOUSH; NASR ESFAHANI MOHAMMAD HOSSEIN; NOURMOHAMMADI ZAHRA; KHAZAIE YAHYA; FORUZANFAR MAHBOUBEH; KIANI ABBAS

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Journal: GENETICS IN THE 3RD MILLENNIUM Year:2011 | Volume:9 | Issue:1 Page(s): 2267-2275

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Title

CONSTRUCTION OF MOUSE PEP CDNA AND EGFP IN TET OFF EXPRESSION VECTOR

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Keywords

PEROXISOMEQ2

PEROXISOMAL PROTEINQ2

EXPRESSION SYSTEMQ2

Abstract

PEROXISOMEs are regarded as one of the organelles in eukaryotic cells with various functions. One of the matrix proteins for PEROXISOME is termed PEROXISOMAL PROTEIN (PEP) which was formerly cloned in mice in 2002. PEP consists of 209 amino acid residues and seems to have a possible role in tissue differentiation of mouse embryo. Studies have shown that PEP expression is increased during myogenesis and neurogenesis. To study the further role of PEP in neurogenesis, it was required to construct an expression vector of this gene to be regulated using Doxycycline or tetracycline in different stages of neurogenesis. Thus, appropriated primers were designed introducing restriction enzymes at the head and tail of the fragment. Moreover, Kozak sequences were considered for high expression of PEP protein. PEP cDNA was amplified and ligated into pTRE2pur vector of Tet off EXPRESSION SYSTEM. Tet off EXPRESSION SYSTEM contains two vectors, a vector called Tet off which produces a protein element which controls the gene expression in the second vector called pTRE2pur off via tetracycline or Doxycycline induction. Tet off vectors can be transferred into cells. In addition to generating TRE2pur/PEP, we inserted EGFP cDNA into the vector TRE2pur and made TRE2pur/EGFP structure to assess the functionality of this system in the cells.

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APA: Copy

MONTAZERI NAJAFABADI, BEHSHAD, GHAEDI, KAMRAN, DORMANI, KIANOUSH, NASR ESFAHANI, MOHAMMAD HOSSEIN, NOURMOHAMMADI, ZAHRA, KHAZAIE, YAHYA, FORUZANFAR, MAHBOUBEH, & KIANI, ABBAS. (2011). CONSTRUCTION OF MOUSE PEP CDNA AND EGFP IN TET OFF EXPRESSION VECTOR. GENETICS IN THE 3RD MILLENNIUM, 9(1), 2267-2275. SID. https://sid.ir/paper/117197/en

Vancouver: Copy

MONTAZERI NAJAFABADI BEHSHAD, GHAEDI KAMRAN, DORMANI KIANOUSH, NASR ESFAHANI MOHAMMAD HOSSEIN, NOURMOHAMMADI ZAHRA, KHAZAIE YAHYA, FORUZANFAR MAHBOUBEH, KIANI ABBAS. CONSTRUCTION OF MOUSE PEP CDNA AND EGFP IN TET OFF EXPRESSION VECTOR. GENETICS IN THE 3RD MILLENNIUM[Internet]. 2011;9(1):2267-2275. Available from: https://sid.ir/paper/117197/en

IEEE: Copy

BEHSHAD MONTAZERI NAJAFABADI, KAMRAN GHAEDI, KIANOUSH DORMANI, MOHAMMAD HOSSEIN NASR ESFAHANI, ZAHRA NOURMOHAMMADI, YAHYA KHAZAIE, MAHBOUBEH FORUZANFAR, and ABBAS KIANI, “CONSTRUCTION OF MOUSE PEP CDNA AND EGFP IN TET OFF EXPRESSION VECTOR,” GENETICS IN THE 3RD MILLENNIUM, vol. 9, no. 1, pp. 2267–2275, 2011, [Online]. Available: https://sid.ir/paper/117197/en

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