P-388 cpDNA KpnI-PstI (0.9 kb) psbA mutated

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Boynton-Gillham laboratory, Duke University, 1991

Chloroplast DNA KpnI-PstI (0.9 kb) in KS+; psbA exon 5, mutated to stop codon at aa 345

host strain: JM109
amp resistant

 

More information

Insert: KpnI-PstI (0.9 kb)

Genes present: psbA exon 5

Vector: KS+

Bacterial host strain: JM109

Selectable marker: amp-r

Origin: Amnon Lers in Boynton/Gillham lab, 1991

P-388

The KS+ vector was digested with KpnI + PstI and a Kpn-Pst insert of 0.9 kb which includes exon 5 of psbA was inserted. This exon was mutated in vitro to change the aa codon ser-345 to a TAA stop codon followed by another change to create a new AflII restriction site. The oligo used in this mutagenesis was psbAstop. This construct was used to obtain the NP mutant.

Lers A, Heifetz PB, Boynton JE, Gillham NW, Osmond CB (1992) The carboxyl-terminal extension of the D1 protein of photosystem II is not required for optimal photosynthetic performance under CO2- and light-saturated growth conditions. J Biol Chem 267:17494-17497