P-240 cpDNA KpnI (3.0 kb) C. smithii spr

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Boynton-Gillham laboratory, Duke University, 1990

Chloroplast DNA KpnI (3.0 kb) in pUC18; 16S rRNA gene from C. smithii spectinomycin resistant mutant. The DNA in this strain came from CC-2477, which was a spectinomycin-resistant isolate from the C. smithii/C. reinhardtii hybrid strain CC-1852. The chloroplast genome in this strain was derived from C. smithii.

host strain: JM83
amp resistant

 

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Strain: CC-2477 C. smithii / C. reinhardtii hybrid

Insert: KpnI (3.0 kb)

Genes present: 16S rRNA

Vector: pUC18

Bacterial host strain: JM83

Selectable marker: amp-r, white on X-gal

Origin: Scott Newman in Boynton/Gillham lab, 1990

P-240

CC-2477 is a spectinomycin-resistant derivative of CC-1852, a hybrid between C. smithii (CC-1373) and C. reinhardtii (CC-124). The CC-124 parent was treated with acriflavine to eliminate mitochondrial DNA before mating, so CC-1852 and its derivatives contain the C. smithii versions of both chloroplast and mitochondrial DNA in a hybrid nuclear background. See Boynton et al. (1987) Proc. Natl. Acad. Sci. USA 84, 2391-2395 for derivation of the strain, and Newman et al. (1990) Genetics 126, 875-888 for use of CC-2477 in subsequent work.

The spectinomycin resistance mutation is an A-> G change at position 1123 in the C. reinhardtii 16S rRNA gene which confers high level spectinomycin resistance and destroys an AatII restriction site.