Expression of the arylsulfatase gene from the beta 2-tubulin promoter in Chlamydomonas reinhardtii.
Arylsulfatase, produced by Chlamydomonas reinhardtii during sulfur-limited growth, is secreted into the periplasmic space and is readily assayed using a chromogenic substrate. To assess the usefulness of the gene encoding arylsulfatase (ars) as a reporter gene in C. reinhardtii, we have fused the promoter region of the beta 2-tubulin gene (tubB2) to the coding region of an ars genomic clone to form a tubB2/ars chimeric sequence. This construct was introduced into C. reinhardtii, strain CC425 (cw-15, arg-2), via cotransformation with the argininosuccinate lyase gene (which complements the arg-2 lesion) (1). Transformants expressing arylsulfatase (Ars) in sulfur-sufficient medium were isolated and subsequently shown to contain the tubB2/ars gene. RNA analysis determined that tubB2/ars transcripts accumulated in these cells. Abundance of the chimeric transcript increased immediately following deflagellation in a manner similar to that of the endogenous tubB2 transcript. Thus, chimeric genes incorporating ars coding sequences and heterologous promoters can be used to examine regulated gene expression in C. reinhardtii.
Davies
J P
JP
Carnegie Institution of Washington, Department of Plant Biology, Stanford, CA 94305.
Weeks
D P
DP
Grossman
A R
AR
D000818:Animals; D001192:Arylsulfatases / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D016825:Chlamydomonas reinhardtii / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D005786:Gene Expression Regulation / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D014404:Tubulin / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D014404:Tubulin; D001192:Arylsulfatases