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首页> 外文期刊>Plant physiology >INTRACELLULAR CARBONIC ANHYDRASE IS ESSENTIAL TO PHOTOSYNTHESIS IN CHLAMYDOMONAS REINHARDTII AT ATMOSPHERIC LEVELS OF CO2 - DEMONSTRATION VIA GENOMIC COMPLEMENTATION OF THE HIGH-CO2-REQUIRING MUTANT CA-1
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INTRACELLULAR CARBONIC ANHYDRASE IS ESSENTIAL TO PHOTOSYNTHESIS IN CHLAMYDOMONAS REINHARDTII AT ATMOSPHERIC LEVELS OF CO2 - DEMONSTRATION VIA GENOMIC COMPLEMENTATION OF THE HIGH-CO2-REQUIRING MUTANT CA-1

机译：胞内碳酸酐酶对大气中CO2含量的莱茵衣原体的光合作用至关重要-通过遗传补充高CO2突变型CA-1进行示范

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Genomic complementation of the high-CO2-requiring mutant ca-1-12-1C of Chlamydomonas reinhardtii was achieved by transformation with DNA pools from an indexed cosmid library of wildtype genomic DNA. Transformation of mutant cells with cosmid DNA from two microtiter plates in the library produced colonies that grew phototrophically at atmospheric CO2 levels. Transformations with cosmid DNA from each of the rows and files of the two plates pinpointed one well in each plate with a cosmid bearing the targeted gene. Sequencing of cosmid subclones revealed a gene encoding a recently identified C. reinhardtii chloroplast carbonic anhydrase (CAH3). Transformations with chimeric constructs combining different portions of the wild-type and mutant genes indicated the presence of a mutation in the 5'-half of the gene. Comparison of mutant and wild-type gene sequences in this region revealed a C-to-A substitution in the mutant gene, which produced a nonsense codon. The data presented demonstrate that the carbonic anhydrase produced from the CAH3 gene is essential to the inorganic carbon-concentrating mechanism in C. reinhardtii and that genomic complementation can be a facile and efficient means for isolating genes associated with defects affecting photosynthesis and other physiological processes in this eukaryotic green alga. [References: 42]
机译：通过用野生型基因组DNA的索引粘粒文库中的DNA库进行转化，可实现莱茵衣藻高CO2突变型ca-1-12-1C的基因组互补。来自库中两个微量滴定板的粘粒DNA突变细胞的转化产生了菌落，该菌落在大气CO2水平下光养生长。来自两个板的每一行和每个文件的粘粒DNA的转化用带有靶向基因的粘粒在每个板中精确定位一个孔。粘粒亚克隆的测序揭示了一个编码最近鉴定的莱茵衣藻叶绿体碳酸酐酶（CAH3）的基因。用结合了野生型和突变基因不同部分的嵌合构建体进行转化表明该基因的5'部分存在突变。比较该区域中的突变型和野生型基因序列，发现突变型基因中的C到A取代产生了无意义的密码子。所提供的数据表明，从CAH3基因产生的碳酸酐酶对于莱茵衣藻中的无机碳浓缩机制必不可少，并且基因组互补可以是分离与影响光合作用和其他生理过程的缺陷相关的基因的简便有效方法。这种真核绿色藻类。 [参考：42]

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