基于PCR-RFLP法鉴定MTHFR基因A1298C位点的内切酶的筛选
Screening of Endonu-cleases for Identification of A1298C Locus of MTHFR Gene by PCR-RFLP

DOI: 10.12677/HJCB.2019.92002, PP. 9-13

Keywords: MTHFR基因，A1298C位点，PCR-RFLP，创造酶切位点
Methylenetetrahydrofolate Reductase Gene, A1298C Locus, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism, Created Restriction Site

Full-Text   Cite this paper   Add to My Lib

Abstract:

寻找价格适宜的限制性内切酶，以期用于PCR-RFLP法对MTHFR基因A1298C位点的分型。根据dbSNP数据库中MTHFR基因A1298C位点的序列，通过各种在线酶切位点分析工具，查找可鉴定该位点的酶，再通过价格对比和分析多态位点上、下游是否存在干扰序列，从而确定候选酶。结合PCR创造酶切位点技术，一种价格便宜的常用酶HinfI具有鉴别该位点的潜力，不仅为该位点的独立检测并且为其与MTHFR基因C677T位点的联合检测奠定了基础。
The aim is to search restrictive endonuclease with suitable price for the genotyping of A1298C locus of MTHFR gene by PCR-RFLP. According to the sequence of A1298C locus of MTHFR gene in dbSNP database, various on-line restriction site analysis tools were used to select the enzymes that could identify the site, and then through price comparison and analysis of whether there were interference sequences in the upstream and downstream of the polymorphic locus, the candidate enzymes were determined. A cheap enzyme, HinfI, has the potential to identify A1298C locus by PCR primer introduced restriction analysis, which laid a foundation for the joint detection of A1298C locus and C677T locus of MTHFR gene.

References

[1]	Du, B., Tian, H., Tian, D., Zhang, C., Wang, W., Wang, L., Ge, M., Hou, Q. and Zhang, W. (2018) Genetic Polymor-phisms of Key Enzymes in Folate Metabolism Affect the Efficacy of Folate Therapy in Patients with Hyperhomocysteinaemia. The British Journal of Nutrition, 119, 887-895. https://doi.org/10.1017/S0007114518000508
[2]	Fekih Mrissa, N., Mrad, M., Klai, S., Zaouali, J., Sayeh, A., Mazigh, C., Nsiri, B., Machgoul, S., Gritli, N. and Mrissa, R. (2013) Association of Methylenetetrahydro Folate Reductase A1298C Polymorphism but Not of C677T with Multiple Sclerosis in Tunisian Patients. Clinical Neurology and Neurosurgery, 115, 1657-1660. https://doi.org/10.1016/j.clineuro.2013.02.025
[3]	Yu, D., Zhuang, Z., Wen, Z., Zang, X. and Mo, X. (2017) MTHFR A1298C Polymorphisms Reduce the Risk of Congenital Heart Defects: A Meta-Analysis from 16 Case-Control Studies. Italian Journal of Pediatrics, 43, 108. https://doi.org/10.1186/s13052-017-0425-1
[4]	Ehsani, M., Imani, A. and Moravveji, A. (2018) Prevalence of Factor V Leiden, MTHFR C677T and MTHFR A1298C Polymorphisms in Patients with Deep Vein Thrombosis in Central Iran. Molecular Biology Reports, 45, 621-624. https://doi.org/10.1007/s11033-018-4201-0
[5]	Allen, R.A., Gatalica, Z., Knezetic, J., Hatcher, L., Vogel, J.S. and Dunn, S.T. (2007) A Common 1317TC Polymorphism in MTHFR Can Lead to Erroneous 1298AC Genotyping by PCR-RE and TaqMan Probe Assays. Genetic Testing, 11, 167-173. https://doi.org/10.1089/gte.2006.0513
[6]	Al-Shahrani, H., Al-Dabbagh, N., Al-Dohayan, N., Arfin, M., Al-Asmari, M., Rizvi, S. and Al-Asmari, A. (2016) Association of the Methylenetetrahydrofolate Reductase (MTHFR) C677T Polymorphism with Primary Glaucoma in Saudi Population. BMC ophthalmology, 16, 156. https://doi.org/10.1186/s12886-016-0337-7

Full-Text