SITE DIRECTED MUTAGENESIS TO PRODUCE MUTANT IRANIAN RICE-NSLTP2 (ORIZA SATIVA) PROTEIN THAT CONTAINS Y45WMUTATION

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AHMADI AZAM; MIROLIAEI MEHRAN; MOTEVALIBASHI MAJID; SADEGHI ABDOLRAHIM; ABTAHI HAMID; ESLAMI ZAHRA

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Journal: Journal of Molecular and Cellular Researches Year:2015 | Volume:28 | Issue:1 Page(s): 1-8

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Title

SITE DIRECTED MUTAGENESIS TO PRODUCE MUTANT IRANIAN RICE-NSLTP2 (ORIZA SATIVA) PROTEIN THAT CONTAINS Y45WMUTATION

Author(s)

Keywords

NSLTP2Q2

MUTAGENESISQ2

SOE-PCRQ2

DRUG DELIVERY SYSTEMQ2

Abstract

NsLTPs (nonspecific lipid transfer proteins) are plant proteins subdivided into nsLTP1(9kDa) and NSLTP2 (7kDa) according to the molecular weight. These highly stable proteins can protect drugs against oxidation or degradations. Rice NSLTP2 due to the inherent ability to cross biological membranes and bind to steroid compounds (including some pharmaceutical compounds) have potential application for use in DRUG DELIVERY SYSTEMs. To enhance the biological capability of this protein in this system, structural alterations are indispensable. These alterations can be created by desired mutations in the encoding genes. Analysis by different software showed that the hydrophobic cavity of the active site plays a prominent role. After designing of mutagenic primers (Tyrosine 45 to Tryptophan (Y45W)) the method of SOE-PCR was used to perform site directed MUTAGENESIS in the gene of rice NSLTP2. Wild type NSLTP2 was cloned into the PGEX6p2 vector. This wild type plasmid was used as a template for constructing the mutant fragment and designed mutant primers are used for amplification of mutant fragments. Mutant rice NSLTP2 are expressed in the pET32a vector in to the E. coli BL21 (DE3) strain. After the amplification of the mutant fragments and confirmation of mutations generated by sequencing, the mutant constructs cloned into the pET32a vector and the fusion mutant protein with a His-tag was purified by Ni-NTA affinity chromatography. The purpose was to create mutations with increased florescence signals. We hope that, in the future, these findings would be helpful in creating biosensors or as drug carrier vehicles.

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APA: Copy

AHMADI, AZAM, MIROLIAEI, MEHRAN, MOTEVALIBASHI, MAJID, SADEGHI, ABDOLRAHIM, ABTAHI, HAMID, & ESLAMI, ZAHRA. (2015). SITE DIRECTED MUTAGENESIS TO PRODUCE MUTANT IRANIAN RICE-NSLTP2 (ORIZA SATIVA) PROTEIN THAT CONTAINS Y45WMUTATION. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 28(1), 1-8. SID. https://sid.ir/paper/248488/en

Vancouver: Copy

AHMADI AZAM, MIROLIAEI MEHRAN, MOTEVALIBASHI MAJID, SADEGHI ABDOLRAHIM, ABTAHI HAMID, ESLAMI ZAHRA. SITE DIRECTED MUTAGENESIS TO PRODUCE MUTANT IRANIAN RICE-NSLTP2 (ORIZA SATIVA) PROTEIN THAT CONTAINS Y45WMUTATION. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2015;28(1):1-8. Available from: https://sid.ir/paper/248488/en

IEEE: Copy

AZAM AHMADI, MEHRAN MIROLIAEI, MAJID MOTEVALIBASHI, ABDOLRAHIM SADEGHI, HAMID ABTAHI, and ZAHRA ESLAMI, “SITE DIRECTED MUTAGENESIS TO PRODUCE MUTANT IRANIAN RICE-NSLTP2 (ORIZA SATIVA) PROTEIN THAT CONTAINS Y45WMUTATION,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 28, no. 1, pp. 1–8, 2015, [Online]. Available: https://sid.ir/paper/248488/en

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