Volume 9, Issue 2 (2018)
JMBS 2018, 9(2): 277-284 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
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 ,
Abstract: (6162 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
References
1. Dini Torkamani MR, Carapetian J. An investigation of physical and chemical characteristics of seed in ten sesame (Sesamum indicum L.) varieties. Iran J Biol. 2008;20(4):327-33. [Persian] [Link]
2. Malek F. Oilseeds & vegetable oils. Tehran: Publishing and Promoting Agricultural Training; 2010. [Persian] [Link]
3. Mosallaieepour Yazdi M, Eghtesadi Sh, Kaseb F, Afkhami Ardakani M, Hoseini F. Effects of sesame oil on blood glucose and lipid profile in type II diabetic patients referring to the Yazd diabetes research center. J Shahid Sadoughi Univ Med Sci. 2008;16(2):15-23. [Link]
4. Fraser CM, Chapple C. The phenylpropanoid pathway in Arabidopsis. Arabidopsis Book. 2011;9:e0152. [Link] [DOI:10.1199/tab.0152]
5. Mc Cann MJ, Gill CI, Mc Glynn H, Rowland IR. Role of mammalian lignans in the prevention and treatment of prostate cancer. Nutr Cancer. 2005;52(1):1-14. [Link] [DOI:10.1207/s15327914nc5201_1]
6. Ono E, Nakai M, Fukui Y, Tomimori N, Fukuchi-Mizutani M, Saito M, et al. Formation of two methylenedioxy bridges by a Sesamum CYP81Q protein yielding a furofuran lignan, (+)-sesamin. Proc Natl Acad Sci U S A. 2006;103(26):10116-21. [Link] [DOI:10.1073/pnas.0603865103]
7. Hukkanen J, Pelkonen O, Hakkola J, Raunio H. Expression and regulation of xenobiotic-metabolizing cytochrome P450 (CYP) enzymes in human lung. Crit Rev Toxicol. 2002;32(5):391-411. [Link] [DOI:10.1080/20024091064273]
8. Hirose N, Inoue T, Nishihara K, Sugano M, Akimoto K, Shimizu S, et al. Inhibition of cholesterol absorption and synthesis in rats by sesamin. J Lipid Res. 1991;32(4):629-38. [Link]
9. Akimoto K, Kitagawa Y, Akamatsu T, Hirose N, Sugano M, Shimizu S, et al. Protective effects of sesamin against liver damage caused by alcohol or carbon tetrachloride in rodents. Ann Nutr Metab. 1993;37(4):218-24. [Link] [DOI:10.1159/000177771]
10. Kuo PC, Lin MC, Chen GF, Yiu TJ, Tzen JT. Identification of methanol-soluble compounds in sesame and evaluation of antioxidant potential of its lignans. J Agric Food Chem. 2011;59(7):3214-9. [Link] [DOI:10.1021/jf104311g]
11. Hata N, Hayashi Y, Okazawa A, Ono E, Satake H, Kobayashi A. Effect of photoperiod on growth of the plants, and sesamin content and CYP81Q1 gene expression in the leaves of sesame (Sesamum indicum L.). Environ Exp Bot. 2012;75:212-9. [Link] [DOI:10.1016/j.envexpbot.2011.07.004]
12. Dar AA, Arumugam N. Lignans of sesame: Purification methods, biological activities and biosynthesis--a review. Bioorg Chem. 2013;50:1-10. [Link] [DOI:10.1016/j.bioorg.2013.06.009]
13. Truan JS, Chen JM, Thompson LU. Comparative effects of sesame seed lignan and flaxseed lignan in reducing the growth of human breast tumors (MCF-7) at high levels of circulating estrogen in athymic mice. Nutr Cancer. 2012;64(1):65-71. [Link] [DOI:10.1080/01635581.2012.630165]
14. Shoun H, Tanimoto T. Denitrification by the fungus Fusarium oxysporum and involvement of cytochrome P-450 in the respiratory nitrite reduction. J Biol Chem. 1991;266(17):11078-82. [Link]
15. Sambrook J, Russel DW. Molecular cloning: A laboratory manual. Long Island: CSHL Press; 2001. [Link]
16. Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, et al. Protein identification and analysis tools on the ExPASy server. In: Walker JM, editor. The proteomics protocols handbook. Heidelberg: Springer Science & Business Media; 2005. pp. 571-607. [Link] [DOI:10.1385/1-59259-890-0:571]
17. Petersen TN, Brunak S, Von Heijne G, Nielsen H. SignalP 4.0: Discriminating signal peptides from transmembrane regions. Nat Methods. 2011;8(10):785-6. [Link] [DOI:10.1038/nmeth.1701]
18. Kato MJ, Chu A, Davin LB, Lewis NG. Biosynthesis of antioxidant lignans in Sesamum indicum seeds. Phytochemistry. 1998;47(4):583-91. [Link] [DOI:10.1016/S0031-9422(97)00727-9]
19. Jiao Y, Davin LB, Lewis NG. Furanofuran lignan metabolism as a function of seed maturation in Sesamum indicum: Methylenedioxy bridge formation. Phytochemistry. 1998;49(2):387-94. [Link] [DOI:10.1016/S0031-9422(98)00268-4]
20. Kim HJ, Ono E, Morimoto K, Yamagaki T, Okazawa A, Kobayashi A, et al. Metabolic engineering of lignan biosynthesis in Forsythia cell culture. Plant Cell Physiol. 2009;50(12):2200-9. [Link] [DOI:10.1093/pcp/pcp156]
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |