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！

丹参果糖磷酸酶基因的克隆和功能研究

谭何新,  刘颖,  张磊

downloadPDF
文章导航 > 药学实践与服务  > 2014 >  32(1): 35-41,48
谭何新, 刘颖, 张磊. 丹参果糖磷酸酶基因的克隆和功能研究[J]. 药学实践与服务, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
引用本文: 谭何新, 刘颖, 张磊. 丹参果糖磷酸酶基因的克隆和功能研究[J]. 药学实践与服务, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
shu
Tan Hexin, Liu Ying, Zhang Lei. Cloning and molecular characterization of fructose-1, 6-bisphosphate aldolase gene from Salvia miltiorrhiza[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
Citation: Tan Hexin, Liu Ying, Zhang Lei. Cloning and molecular characterization of fructose-1, 6-bisphosphate aldolase gene from Salvia miltiorrhiza[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
shu

丹参果糖磷酸酶基因的克隆和功能研究

doi: 10.3969/j.issn.1006-0111.2014.01.009

  • 第二军医大学药学院药用植物学教研室, 上海 200433

基金项目: 国家自然科学基金项目(31160059);上海市科委浦江人才项目(13PJ1411000).

Cloning and molecular characterization of fructose-1, 6-bisphosphate aldolase gene from Salvia miltiorrhiza

  • Department of Pharmaceutical Botany, School of Pharmacy, Second Military Medical University, Shanghai 200433, China

  • 摘要: 丹参是著名的药用植物,在中国、日本、美国和欧洲国家被广泛地用于心脑血管系统疾病的治疗。笔者首次从丹参中克隆出了一个新的果糖磷酸酶基因,并将其命名为SmFBA,GenBank编号为FJ540907。SmFBA的cDNA全长含有1 390个核苷酸,包含一个完整的1 065 bp的开放阅读框,编码355个氨基酸残基。SmFBA基因全长含有3个外显子和2个内含子。生物信息学分析显示SmFBA蛋白预测的等电点为5.60,预测的分子量为37.78 ku,和其他植物物种中果糖磷酸酶具有很高的序列同源性。用Southern杂交技术显示SmFBA在丹参基因组中呈低拷贝。该基因在丹参根、茎、叶等器官都表达,根中表达量最高。体外重组表达的SmFBA在大肠杆菌中具有酶活性,并且能够提高大肠杆菌的耐盐性。这项研究进一步拓展了人们对高等植物糖酵解途径的认识。
    Abstract: Salvia miltiorrhiza (Bung), a famous medicinal plant, is widely used in China, Japan, America and European countries to treat various conditions, such as cardiovascular and cerebrovascular disease, due to their excellent medicinal values. A novel fructose-bisphosphate aldolase gene (designated as SmFBA, GenBank accession no. FJ540907) was cloned from S. miltiorrhiza for the first time. The full-length cDNA of SmFBA was 1 390 bp with an open reading frame (ORF) of 1 065 bp which encoding a protein of 355 amino acid residues. The deduced protein had isoelectric point (pI) of 5.60 and a calculated molecular weight of about 37.78 ku. The deduced amino acid sequence of SmFBA gene shared high homology and identity with other plant FBAs. The SmFBA genomic DNA sequence was also obtained, revealing SmFBA had three exons and two introns. Southern-blot analysis indicated that SmFBA was a low-copy gene in S. miltiorrhiza genomic. Real-time quantitative PCR analysis showed that SmFBA expressed constitutively in all tested organs, with the highest expression level in roots. In addition, the recombinant SmFBA protein has enzyme activity in E. coli and could improve the high-salinity stress tolerance of E. coli. The successful isolation of the SmFBA gene will be helpful for studying EMP pathway in the near future.
  • [1] Siddiqui KA, Banerjee AK. Fructose 1,6-bisphosphate aldolase activity of Rhizobium species[J]. Folia Microbiol,1975, 20:412.
    [2] Fridlyand LE, Scheibe R. Regulation of the Calvin cycle for CO2 fixation as an example for general control mechanisms in metabolic cycles[J]. BioSystems,1999, 51:79.
    [3] Jones JB. Enzymes in organic synthesis[J]. Tetrahedron Lett,1986, 42:3351.
    [4] Rutter WJ. Evolution of aldolase[J]. Fed Proc,1964, 23:1248.
    [5] Marsh JJ, Lebherz HG. Fructose-bisphosphate aldolases:an evolutionary history[J]. Trends Biochem Sci,1992, 17:110.
    [6] Doolittle RF. Convergent evolution:the need to be explicit[J]. Trends Biochem Sci,1994, 19:15.
    [7] Perham RN. The fructose 1,6-bisphosphate aldolases:same reaction, different enzymes[J]. Biochem Soc Transcript,1990, 18:185.
    [8] Siebers B, Brinkmann H, Dorr C, et al. Archaeal fructose-1,6-bisphosphate aldolases constitute a new family of archaeal type class I aldolase[J]. J Biol Chem,2001, 276:28710.
    [9] Effenberger F, Straub A. A novel convenient preparation of dihydroxyacetonphosphate and its use in enzymatic aldol reactions[J]. Tetrahedron Lett, 1987, 28:1641.
    [10] Anderson LE, Advani VR. Chloroplast and cytoplasmic enzymes. Three distinct isoenzymes associated with the reductive pentose phosphate cycle[J]. Plant Physiol,1970, 45:583.
    [11] Briggs WR, Christie JM. Phototropins 1 and 2:versatile plant blue-light receptors[J]. Trends Plant Sci,2002, 7:204.
    [12] Joh K, Arai Y, Mukai T, et al. Expression of three mRNA species from a single rat aldolase A gene, differing in their 5' non-coding regions[J]. J Mol Biol,1986, 190:401.
    [13] Maire P, Gauttrons S, Hakim V, et al. Characterization of three optional promoters in the 5'-region of the human aldolase A gene[J]. J Mol Biol,1987, 197:425.
    [14] Kai GY, Jiang JH, Zhao DL, et al. Isolation and expression profile analysis of a new cDNA encoding 5-alpha-taxadienol-10-beta-hydroxylase from Taxus media[J]. J Plant Biochem Biotechnol,2006, 15:1.
    [15] Zhou LM, Zuo Z, Moses S. Danshen:an overview of its chemistry, pharmacology, pharmacokinetics, and clinical use[J]. J Clin Pharmacol,2005, 45:1345.
    [16] Jaakola L, Pirttila AM, Halonen M, et al. Isolation of high quality RNA from bilberry (Vaccinium myrtillus L.) fruit[J]. Mol Biotechnol,2001, 19:201.
    [17] Saitou N, Nei M. The neighbor-joining method:a new method for reconstructing phylogenetic trees[J]. Mol Biol Evol,1987, 4:406.
    [18] Thompson JD, Higgins DG, Gibson TJ. Clustal-W-improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice[J]. Nucl Acids Res,199422:4673.
    [19] Kumar S, Tamura K, Jakobsen IB, et al. MEGA2:molecular evolutionary genetics analysis software[J]. Bioinformatics,2001, 17, 1244.
    [20] Fan W, Zhang Z, Zhang Y. Cloning and molecular characterization of fructose-1,6-bisphosphate aldolase gene regulated by high-salinity and drought in Sesuvium portulacastrum[J]. Plant Cell Rep,2009, 28(6):975.
  • [1] 陈静, 何瑞华, 翁月, 徐熠, 刘静, 黄瑾.  基于网络药理学和分子对接技术探究定清片活性成分治疗白血病的作用机制 . 药学实践与服务, 2024, 42(11): 479-486. doi: 10.12206/j.issn.2097-2024.202401073
    [2] 陈金涛, 乔子婴, 马明华, 张若曦, 王振伟, 年华.  基于网络药理学和分子对接技术研究金芪清疏颗粒治疗社区获得性肺炎的潜在机制 . 药学实践与服务, 2024, 42(11): 471-478. doi: 10.12206/j.issn.2097-2024.202312014
    [3] 姚小静, 计佩影, 陆峰, 施国荣, 傅翔.  表面增强拉曼光谱法快速测定尿液中曲马多的研究 . 药学实践与服务, 2024, 42(): 1-5. doi: 10.12206/j.issn.2097-2024.202401072
    [4] 夏哲炜, 曾垣烨, 朱海菲, 李育, 陈啸飞.  核磁共振磷谱法测定磷酸氢钙咀嚼片中药物含量 . 药学实践与服务, 2024, 42(9): 399-401, 406. doi: 10.12206/j.issn.2097-2024.202404063
    [5] 黄韵, 张正银, 金英, 郑怡菁, 李铁军, 孙莉莉.  耐碳青霉烯类肺炎克雷伯菌及大肠埃希菌临床分离株耐药性及耐药基因分析 . 药学实践与服务, 2024, 42(10): 439-444. doi: 10.12206/j.issn.2097-2024.202309059
    [6] 戴菲菲, 傅翔, 陈琼年, 俞苏纯.  上海某二级医院革兰阴性菌流行特征的回顾性分析 . 药学实践与服务, 2024, 42(): 1-5. doi: 10.12206/j.issn.2097-2024.202305005
    [7] 钱淑雨, 李铁军.  耐碳青霉烯类肠杆菌耐药机制的研究进展 . 药学实践与服务, 2024, 42(10): 419-425. doi: 10.12206/j.issn.2097-2024.202405005
    [8] 张成中, 朱雪艳, 卜其涛, 王宏瑞, 黄宝康.  基于网络药理学与分子对接预测鸡骨草特征图谱研究 . 药学实践与服务, 2024, 42(8): 350-358. doi: 10.12206/j.issn.2097-2024.202303048
    [9] 王耀振, 徐灿, 吕顺莉, 田泾, 张东炜.  钾离子竞争性酸阻滞剂的药学特征研究进展 . 药学实践与服务, 2024, 42(7): 278-284. doi: 10.12206/j.issn.2097-2024.202306040
    [10] 张晶晶, 索丽娜, 郑兆红.  89例细菌性肝脓肿的临床特征及抗感染治疗分析 . 药学实践与服务, 2024, 42(6): 267-272. doi: 10.12206/j.issn.2097-2024.202302039
    [11] 王雪莲, 郑斯莉, 李志勇, 罗亨宇, 缪朝玉.  全身过表达人METRNL基因小鼠模型的构建与验证 . 药学实践与服务, 2024, 42(5): 198-202, 222. doi: 10.12206/j.issn.2097-2024.202311014
    [12] 姚瑞阳, 于海征, 李耀盺, 张磊.  丹参FBXL 基因家族的鉴定和表达模式分析 . 药学实践与服务, 2024, 42(11): 461-470. doi: 10.12206/j.issn.2097-2024.202407034
  • 加载中
WeChat 点击查看大图
计量
  • 文章访问数:  3115
  • HTML全文浏览量:  270
  • PDF下载量:  62
  • 被引次数: 0
出版历程
  • 收稿日期:  2012-12-19
  • 修回日期:  2013-10-29

丹参果糖磷酸酶基因的克隆和功能研究

doi: 10.3969/j.issn.1006-0111.2014.01.009
  • 第二军医大学药学院药用植物学教研室, 上海 200433
    • 基金项目:  国家自然科学基金项目(31160059);上海市科委浦江人才项目(13PJ1411000).
  • 收稿日期: 2012-12-19
  • 修回日期: 2013-10-29

摘要: 丹参是著名的药用植物,在中国、日本、美国和欧洲国家被广泛地用于心脑血管系统疾病的治疗。笔者首次从丹参中克隆出了一个新的果糖磷酸酶基因,并将其命名为SmFBA,GenBank编号为FJ540907。SmFBA的cDNA全长含有1 390个核苷酸,包含一个完整的1 065 bp的开放阅读框,编码355个氨基酸残基。SmFBA基因全长含有3个外显子和2个内含子。生物信息学分析显示SmFBA蛋白预测的等电点为5.60,预测的分子量为37.78 ku,和其他植物物种中果糖磷酸酶具有很高的序列同源性。用Southern杂交技术显示SmFBA在丹参基因组中呈低拷贝。该基因在丹参根、茎、叶等器官都表达,根中表达量最高。体外重组表达的SmFBA在大肠杆菌中具有酶活性,并且能够提高大肠杆菌的耐盐性。这项研究进一步拓展了人们对高等植物糖酵解途径的认识。

English Abstract

谭何新, 刘颖, 张磊. 丹参果糖磷酸酶基因的克隆和功能研究[J]. 药学实践与服务, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
引用本文: 谭何新, 刘颖, 张磊. 丹参果糖磷酸酶基因的克隆和功能研究[J]. 药学实践与服务, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
shu
Tan Hexin, Liu Ying, Zhang Lei. Cloning and molecular characterization of fructose-1, 6-bisphosphate aldolase gene from Salvia miltiorrhiza[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
Citation: Tan Hexin, Liu Ying, Zhang Lei. Cloning and molecular characterization of fructose-1, 6-bisphosphate aldolase gene from Salvia miltiorrhiza[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
shu

    全文HTML

参考文献 (20)

目录

    /

    返回文章
    返回

    版权所有：《药学实践与服务》编辑委员会

    主管、主办: 中国人民解放军海军军医大学地址: 上海市杨浦区国和路325号邮政编码: 200433

    电话: （021）81871324,81871397 E-mail: yxsjzzs@163.com

    网站备案号 沪ICP备05003363号-1

    本系统由 北京仁和汇智信息技术有限公司 开发    百度统计