Volume 9, Issue 2 (2018)
JMBS 2018, 9(2): 277-284 |
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Hemati S, Dehghan Nayeri F. Cloning, Sequencing, and Bioinformatics Study of CYP81Q1 Gene of Iranian Sesame (Seamum indicum L.) Cultivar. JMBS 2018; 9 (2) :277-284
URL: http://biot.modares.ac.ir/article-22-13348-en.html
URL: http://biot.modares.ac.ir/article-22-13348-en.html
1- Agricultural Biotechnology Department, Agricultural & Natural Sciences Faculty, Imam Khomeini International University, Qazvin, Iran
2- Agricultural Biotechnology Department, Agricultural & Natural Sciences Faculty, Imam Khomeini International University, Qazvin, Iran, Agricultural Biotechnology Department, Agricultural & Natural Sciences Faculty, Imam Khomeini International University, Qazvin, Iran. Postal Code: 3414896818 , nayeri@ut.ac.ir
2- Agricultural Biotechnology Department, Agricultural & Natural Sciences Faculty, Imam Khomeini International University, Qazvin, Iran, Agricultural Biotechnology Department, Agricultural & Natural Sciences Faculty, Imam Khomeini International University, Qazvin, Iran. Postal Code: 3414896818 , nayeri@ut.ac.ir
Abstract: (4924 Views)
Aims: Antioxidants in sesame oil, including tocopherols and sesamin have greatly increased the shelf life of it against heat. Following the increase in the expression of the cytochrome P450 enzyme encoder (CYP81Q1), the content of sesame is increased in different stages of development of sesame seeds. The aim of this study was cloning, sequencing, and bioinformatics study of CYP81Q1 gene of Iranian sesame (Seamum indicum L.) cultivar.
Materials and Methods: In the present experimental research, DNA was extracted from leaves and stems of Karaj1 sesame cultivar and the target gene was amplified by PCR. Gene was cloned in binary vector pBI121 and confirmed by 3 methods, including enzymatic digestion, PCR, and sequencing. Then bioinformatics characterization of this gene was studied and the Ramachandran plot was drawn on the three-dimensional structure of the gene.
Findings: Cloning was confirmed. DNA sequencing results confirmed the cloned segment. Molecular weight and predicted isoelectric point of the protein were 57021.3 Dalton and 8.46, respectively. The three-dimensional structure of the protein had a good stroke chain. The sequencing result of this gene showed a difference in the 23 nucleotides of this gene in sesame seeds of Karaj 1 (access number KP771974.1) with a reported sequence in the NCBI gene bank (access number AB194714.1), which resulted in the sequencing of the CYP81Q1 gene in Iranian sesame (Karaj 1) at this database.
Conclusion: Based on nucleotide sequencing, the target gene has 1521 base pairs, and differs from 23 nucleotides with the sample registered at the NCBI World Bank. This gene encodes a protein length of 506 amino acids. This protein is very similar with the registered protein in NCBI.
Materials and Methods: In the present experimental research, DNA was extracted from leaves and stems of Karaj1 sesame cultivar and the target gene was amplified by PCR. Gene was cloned in binary vector pBI121 and confirmed by 3 methods, including enzymatic digestion, PCR, and sequencing. Then bioinformatics characterization of this gene was studied and the Ramachandran plot was drawn on the three-dimensional structure of the gene.
Findings: Cloning was confirmed. DNA sequencing results confirmed the cloned segment. Molecular weight and predicted isoelectric point of the protein were 57021.3 Dalton and 8.46, respectively. The three-dimensional structure of the protein had a good stroke chain. The sequencing result of this gene showed a difference in the 23 nucleotides of this gene in sesame seeds of Karaj 1 (access number KP771974.1) with a reported sequence in the NCBI gene bank (access number AB194714.1), which resulted in the sequencing of the CYP81Q1 gene in Iranian sesame (Karaj 1) at this database.
Conclusion: Based on nucleotide sequencing, the target gene has 1521 base pairs, and differs from 23 nucleotides with the sample registered at the NCBI World Bank. This gene encodes a protein length of 506 amino acids. This protein is very similar with the registered protein in NCBI.
Article Type: Research Paper |
Subject:
Agricultural Biotechnology
Received: 2016/11/7 | Accepted: 2017/12/16 | Published: 2018/06/21
Received: 2016/11/7 | Accepted: 2017/12/16 | Published: 2018/06/21
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