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[采用聚合酶链反应(PCR)获得玄参科全寄生植物地下肉苁蓉的rbcL基因。与自养植物毛地黄和草原山萝花的比较]

[Obtention with polymerase chain reaction (PCR) of gene rbcL in the scrophulariaceae holoparasite Lathraea clandestina L. Comparison with autotroph Digitalis purpurea L. and Melampyrum pratense L].

作者信息

Thalouarn P, Renaudin S

出版信息

C R Acad Sci III. 1991;313(9):381-7.

PMID:1756411
Abstract

Amyloplastid DNA was extracted from the Scrophulariaceae holoparasite Lathraea clandestina L. and then purified. An rbcL gene amplification was performed using polymerase chain reaction. Two regions of well conserved sequences in Tobacco, Spinach and Maize rbcL gene have been used as primers. PCR yields a sequence of about 1,230 base pairs, almost nine tenths of the rbcL coding region. Identical results were obtained with plastid DNAs from Tobacco and two other species of Scrophulariaceae: the non-parasitic Digitalis purpurea L. and the hemiparasite Melampyrum pratense L. PCR products were digested with BamHI restriction enzyme and no changes were shown in the localization of the restriction site whatever the species assayed. Identical restriction patterns were also observed with Tobacco and Digitalis PCR products restricted with PstI and KpnI, whereas Melampyrum and Lathraea exhibited a different restriction pattern with PstI. So, despite slight differences, some analogies between Lathraea and Digitalis or Tobacco gene were evidenced. Cloning and sequencing of these PCR products could give a more accurate response to the following question: to what extent have changes occurred in the rbcL gene in a plant which lacks chlorophyll?

摘要

从玄参科全寄生植物地下齿草(Lathraea clandestina L.)中提取造粉体DNA,然后进行纯化。使用聚合酶链反应对rbcL基因进行扩增。烟草、菠菜和玉米rbcL基因中两个保守性良好的序列区域被用作引物。聚合酶链反应产生了一个约1230个碱基对的序列,几乎是rbcL编码区的十分之九。从烟草以及玄参科的另外两个物种:非寄生植物毛地黄(Digitalis purpurea L.)和半寄生植物草地山萝花(Melampyrum pratense L.)的质体DNA中也获得了相同的结果。用BamHI限制酶消化聚合酶链反应产物,无论检测的是哪个物种,限制位点的定位都没有变化。用PstI和KpnI限制烟草和毛地黄的聚合酶链反应产物时,也观察到了相同的限制模式,而草地山萝花和地下齿草在用PstI限制时表现出不同的限制模式。因此,尽管存在细微差异,但地下齿草与毛地黄或烟草基因之间仍存在一些相似之处。对这些聚合酶链反应产物进行克隆和测序,可以更准确地回答以下问题:在缺乏叶绿素的植物中,rbcL基因发生了多大程度的变化?

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