The argininosuccinate lyase gene of <i>Chlamydomonas reinhardtii</i> : cloning of the cDNA and its characterization as a selectable shuttle marker

Article Article scientifique Article The argininosuccinate lyase gene of <i>Chlamydomonas reinhardtii</i> : cloning of the cDNA and its characterization as a selectable shuttle marker eng 1999 author Auchincloss Damay Andréa 29803 author Loroch Albert Yvan 29367 author Rochaix Jean-David 25289 21-30 10.1007/s004380050937 0026-8925 http://link.springer.com/10.1007/s004380050937 We describe the cDNA sequence for ARG7, the gene that encodes argininosuccinate lyase – a selectable nuclear marker – in Chlamydomonas reinhardtii. The 5′ end of the cDNA contains one more exon and the organisation of the mRNA is different from that predicted from the genomic sequence. When expressed under the control of the endogenous RbcS2 promoter, the 2.22-kb cDNA complements the arg7 mutation as well as the genomic DNA. A linear cDNA fragment lacking promoter sequences is also able to complement, suggesting that it could be used in promoter-trapping experiments. Despite the presence of a sequence encoding a potential chloroplast transit peptide in the cDNA the protein is not targeted to the chloroplast, nor can it complement the arg7 mutation when expressed there. By inserting a T7 bacteriophage promoter into the plasmid, a version of the cDNA which is able to complement both the C. reinhardtii arg7 mutant and the Escherichia coli argH mutant has been created. This modified Arg7 cDNA provides two advantages over the genomic DNA currently in use for gene tagging: it is shorter (6.2 kb versus 11.9 kb for pARG7.8φ3), and the selectable marker used in C. reinhardtii is the same as that used in E. coli, making plasmid rescue of the tag much more likely to succeed. Chlamydomonas Argininosuccinate lyase cDNA Transformation Shuttle vector Plasmid rescue Dewey 570 43 Département de biologie moléculaire 161 false Molecular and General Genetics 261 1 fd19a3ef-4c65-4b85-b892-d760c4aa654a Article (Published version) Public Auchincloss1999_Article_TheArgininosuccinateLyaseGeneO.pdf 252476 application/pdf openAccess Faculté des sciences / Section de biologie / Département de biologie moléculaire

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The argininosuccinate lyase gene of Chlamydomonas reinhardtii : cloning of the cDNA and its characterization as a selectable shuttle marker	Molecular and General Genetics			1999	324	378

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Loroch, Albert Yvan