[26] Isolation and expression of a cDNA coding for aequorin, the Ca2+-activated photoprotein from Aequorea victoria

摘要: Publisher Summary This chapter discusses isolation and expression of a cDNA coding for aequorin, the Ca 2+ -activated photoprotein from Aequorea victoria . Aequorin is bioluminescent protein isolated luminous hydromedusan victoria, which emits blue light upon binding Isolating aequorin was desirable number reasons. (1) It very laborious procedure to collect process so purification would be greatly simplified strain Escherichia coli expressing apoprotein. (2) Such yield unlimited quantities apoaequorin not only biochemical investigations but also use as nonisotopic label immunodiagnostics. An library constructed using RNA circumoral rings jellyfish. The poly (A) + fraction used prepare double-stranded cloned into Pst I-digested pBR322. E. transformants are screened presence an oligonucleotide mixture whose sequence based on amino acid sequence. Six in 6000 comprising bank hybridized mixture. activity demonstrated extract containing one putative cDNAs.