P-229 cpDNA BamHI-HindIII (5.9 kb) spr

$30.00

Boynton-Gillham laboratory, Duke University, 1989

Chloroplast DNA BamHI-HindIII (5.9 kb) in pUC18, from C. smithii; contains 16S through 5′ end of 23S, with spectinomycin resistance mutation near 3′ end of 16S rRNA. 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

 

More information

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

Insert: BamHI-HindIII (5.9 kb)

Genes present: 16S rRNA, 23S rRNA (5’end)

Vector: pUC18

Bacterial host strain: JM83

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

Origin: Scott Newman in Boynton/Gillham lab, 1989

P-229

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 that confers high level spectinomycin resistance and destroys an AatII restriction site.

Newman SM, Boynton JE, Gillham NW, Randolph-Anderson BL, Johnson AM, Harris EH (1990) Transformation of chloroplast ribosomal RNA genes in Chlamydomonas: molecular and genetic characterization of integration events. Genetics 126:875-888