鼠曲草简单重复序列间扩增反应体系的建立及优化

江圣圭; 董志颖; 李卿; 赵英魁; 黄豆豆; 孙连娜

doi:10.3969/j.issn.1006-0111.201909048

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

x 关闭永久关闭

应中央军委要求，2022年9月起，《药学实践杂志》将更名为《药学实践与服务》，双月刊，正文96页；2023年1月起，拟出版月刊，正文64页，数据库收录情况与原《药学实践杂志》相同。欢迎作者踊跃投稿！

鼠曲草简单重复序列间扩增反应体系的建立及优化

江圣圭, 董志颖, 李卿, 赵英魁, 黄豆豆, 孙连娜

文章导航 > 药学实践与服务 > 2020 > 38(1): 42-46,62

江圣圭, 董志颖, 李卿, 赵英魁, 黄豆豆, 孙连娜. 鼠曲草简单重复序列间扩增反应体系的建立及优化[J]. 药学实践与服务, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048

引用本文:

JIANG Shenggui, DONG Zhiying, LI Qing, ZHAO Yingkui, HUANG Doudou, SUN Lianna. Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine[J]. Journal of Pharmaceutical Practice and Service, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048

Citation:

JIANG Shenggui, DONG Zhiying, LI Qing, ZHAO Yingkui, HUANG Doudou, SUN Lianna. Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine[J]. Journal of Pharmaceutical Practice and Service, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048

鼠曲草简单重复序列间扩增反应体系的建立及优化

doi: 10.3969/j.issn.1006-0111.201909048

1.
福建中医药大学药学院, 福建福州 350122
2.
上海中医药大学中药学院, 上海 201203
3.
海军军医大学附属长征医院药学部, 上海 200003
4.
海军军医大学药学院, 上海 200433
5.
福建中医药大学药学院, 福建福州 350122;上海中医药大学中药学院, 上海 201203

基金项目: 国家自然科学基金项目（81373954），上海市科委资助课题项目（13ZR1448800）

Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine

1.
School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China
2.
School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
3.
Department of Pharmacy, Changzheng Hospital, Navy Medical University, Shanghai 200003, China
4.
School of Pharmacy, Navy Medical University, Shanghai 200433, China
5.
School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

摘要: 目的通过建立并优化鼠曲草简单重复序列间扩增（ISSR-PCR）反应体系，为鼠曲草后续遗传多样性研究提供实验依据。方法本研究首先采用单因素实验法和全面实验法优化鼠曲草ISSR-PCR反应体系，然后在最适体系下，分步进行引物及其退火温度的筛选，最后验证该体系的可行性。结果 ISSR-PCR最适反应体系包括10 μl Premix Taq DNA聚合酶、0.3 μmol/L引物、10 ng DNA模板、灭菌水加至20 μl；从100条通用引物中最终筛选出10条引物；验证结果表明该体系稳定性高、重复性好，且选定引物多态性好。结论本研究首次建立了鼠曲草ISSR-PCR扩增体系，并筛选出合适的引物，为鼠曲草后续遗传多样性研究夯实了基础。
- 鼠曲草 /
- 简单重复序列间扩增 /
- 体系优化 /
- 引物筛选
Abstract: Objective To provide the experimental basis for the subsequent genetic diversity research through establishing and optimizing the inter-simple sequence repeat PCR (ISSR-PCR) reaction system of Gnaphalium affine. Methods The single-factor experimental method and full experimental method were used to optimize the ISSR-PCR reaction system of Gnaphalium affine. Under the optimal system, after screening primers and corresponding annealing temperatures, the systematic feasibility was verified. Results The optimal ISSR-PCR reaction system was consisted of 10 μl Premix Taq DNA polymerase, 0.3 μmol/L primer, 10 ng DNA template, and sterilized water added to 20 μl. Finally, 10 primers were screened from 100 universal primers, and verification results indicated the system had high stability, good reproducibility, and the selected primers had good polymorphism. Conclusion The ISSR-PCR amplification system of Gnaphalium affine was established for the first time and the primers with appropriate annealing temperatures were filtered out, which provided a reference for the subsequent genetic diversity research of Gnaphalium affine.
- Gnaphalium affine /
- inter-simple sequence repeat PCR /
- system optimization /
- primer screening

[1]	LI J L, HUANG D D, CHEN W S, et al. Two new phenolic glycosides from Gnaphalium affine D. Don and their anti-complementary activity[J]. Molecules,2013,18(7):7751-7760
[2]	ZHANG H J, LI L N, ZHOU J, et al. Effects of Gnaphalium affine D. Don on hyperuricemia and acute gouty arthritis[J]. J Ethnopharmacol,2017,203:304-311
[3]	LIN Y, LIU P G, LIANG W Q, et al. Luteolin-4'-O-glucoside and its aglycone, two major flavones of Gnaphalium affine D. Don, resist hyperuricemia and acute gouty arthritis activity in animal models[J]. Phytomedicine,2018,41:54-61
[4]	SEONG Y A, HWANG D, KIM G D. The anti-inflammatory effect of Gnaphalium affine through inhibition of NF-κB and MAPK in lipopolysaccharide-stimulated RAW_264.7 cells and analysis of its phytochemical components[J]. Cell Biochem Biophys,2016,74(3):407-417
[5]	LIN W Q, XIE J X, WU X M, et al. Inhibition of xanthine oxidase activity by Gnaphalium affine extract[J]. Chin Med Sci J,2014,29(4):225-230
[6]	ZHANG W, WU C Z, FAN S Y. Chemical constituents from Gnaphalium affine and their xanthine oxidase inhibitory activity[J]. Chin J Nat Med,2018,16(5):347-353
[7]	高俊斌, 王璇, 陈燕红, 等. HPLC法同时测定鼠曲草中7种成分[J]. 中成药, 2018, 40(5):1116-1119
[8]	GOULÃO L, OLIVEIRA C M. Molecular characterisation of cultivars of apple (Malus×domestica Borkh.) using microsatellite (SSR and ISSR) markers[J]. Euphytica,2001,122(1):81-89
[9]	ZIETKIEWICZ E, RAFALSKI A, LABUDA D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification[J]. Genomics,1994,20(2):176-183
[10]	任梦云, 陈彦君, 张盾, 等. 山莨菪ISSR-PCR反应体系的建立与优化[J]. 分子植物育种, 2017, 15(12):5006-5014
[11]	任风鸣, 胡开治, 刘燕琴, 等. 传统中药金钱草ISSR-PCR反应体系的正交优化研究[J]. 中国中药杂志, 2014, 39(12):2233-2238
[12]	唐辉, 陈宗游, 史艳财, 等. 正交设计优化地枫皮ISSR-PCR反应体系[J]. 中草药, 2013, 44(5):610-615
[13]	张福生, 郭顺星. 金线莲ISSR反应体系的建立与优化[J]. 中草药, 2011, 42(1):137-142
[14]	穆立蔷, 刘赢男, 冯富娟, 等. 紫椴ISSR-PCR反应体系的建立与优化[J]. 林业科学, 2006, 42(6):26-31
[15]	CUI C J, LI Y, LIU Y L, et al. Determination of genetic diversity among Saccharina germplasm using ISSR and RAPD markers[J]. C R Biol,2017,340(2):76-86
[16]	WU W F, CHEN F X, YEH K, et al. ISSR analysis of genetic diversity and structure of plum varieties cultivated in southern China[J]. Biology (Basel),2018,8(1):E2
[17]	刘蕤, 杨际双. 菊属11个野生种和12个栽培品种遗传关系的ISSR分析[J]. 基因组学与应用生物学, 2009, 28(5):874-882

点击查看大图

计量

文章访问数: 4366
HTML全文浏览量: 392
PDF下载量: 3295
被引次数: 0

鼠曲草简单重复序列间扩增反应体系的建立及优化

doi: 10.3969/j.issn.1006-0111.201909048

1. 福建中医药大学药学院, 福建福州 350122

2. 上海中医药大学中药学院, 上海 201203

3. 海军军医大学附属长征医院药学部, 上海 200003

4. 海军军医大学药学院, 上海 200433

5. 福建中医药大学药学院, 福建福州 350122;上海中医药大学中药学院, 上海 201203

基金项目: 国家自然科学基金项目（81373954），上海市科委资助课题项目（13ZR1448800）

收稿日期: 2019-09-12
修回日期: 2019-10-21

关键词:

鼠曲草 /

简单重复序列间扩增 /

体系优化 /

引物筛选

摘要: 目的通过建立并优化鼠曲草简单重复序列间扩增（ISSR-PCR）反应体系，为鼠曲草后续遗传多样性研究提供实验依据。方法本研究首先采用单因素实验法和全面实验法优化鼠曲草ISSR-PCR反应体系，然后在最适体系下，分步进行引物及其退火温度的筛选，最后验证该体系的可行性。结果 ISSR-PCR最适反应体系包括10 μl Premix Taq DNA聚合酶、0.3 μmol/L引物、10 ng DNA模板、灭菌水加至20 μl；从100条通用引物中最终筛选出10条引物；验证结果表明该体系稳定性高、重复性好，且选定引物多态性好。结论本研究首次建立了鼠曲草ISSR-PCR扩增体系，并筛选出合适的引物，为鼠曲草后续遗传多样性研究夯实了基础。

English Abstract

Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine

1. School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China

2. School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

3. Department of Pharmacy, Changzheng Hospital, Navy Medical University, Shanghai 200003, China

4. School of Pharmacy, Navy Medical University, Shanghai 200433, China

5. School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

Received Date: 2019-09-12
Rev Recd Date: 2019-10-21

Keywords:

Abstract: Objective To provide the experimental basis for the subsequent genetic diversity research through establishing and optimizing the inter-simple sequence repeat PCR (ISSR-PCR) reaction system of Gnaphalium affine. Methods The single-factor experimental method and full experimental method were used to optimize the ISSR-PCR reaction system of Gnaphalium affine. Under the optimal system, after screening primers and corresponding annealing temperatures, the systematic feasibility was verified. Results The optimal ISSR-PCR reaction system was consisted of 10 μl Premix Taq DNA polymerase, 0.3 μmol/L primer, 10 ng DNA template, and sterilized water added to 20 μl. Finally, 10 primers were screened from 100 universal primers, and verification results indicated the system had high stability, good reproducibility, and the selected primers had good polymorphism. Conclusion The ISSR-PCR amplification system of Gnaphalium affine was established for the first time and the primers with appropriate annealing temperatures were filtered out, which provided a reference for the subsequent genetic diversity research of Gnaphalium affine.