Cloning of a cDNA encoding a new ribosome-inactivating protein from Beta vulgaris vulgaris (mangold).

By means of a lambda ZAP II cDNA library constructed from seedings of Beta vulgaris vulgaris and immunoscreening, a cDNA clone containing a partial sequence of a new ribosome-inactivating protein (RIP) was obtained. As confirmed by Western blot analysis, this clone produced a RIP upon induction with IPTG. We called it betavulgin (Bvg). The recombinant protein (re-Bvg) was somewhat smaller than plant-derived RIP (28 versus 30 and 32 kDa), but showed the specific N-glycosidase activity on tobacco ribosomes, confirming its RIP character. The cDNA was sequenced and the missing 5'-end was established by RACE using bvg-specific primers. The entire cDNA was 1080 nucleotides in length and encoded a protein of 272 amino acids with a sequence identity of 26-40% with other RIP.