Cloning and expression of a gene encoding gelonin, a ribosome-inactivating protein from Gelonium multiflorum

• Published in: Gene Vol. 134; no. 2; pp. 223 - 227
• Main Authors: Nolan, Patricia A., Garrison, Darryl A., Better, Marc
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 08.12.1993; Amsterdam Elsevier; New York, NY
• Subjects: Amino Acid Sequence; amino acid sequences; Base Sequence; Biological and medical sciences; Cloning, Molecular; complementary DNA; DNA; Escherichia coli; Fundamental and applied biological sciences. Psychology; gel gene; genbank/l12243; Genes. Genome; Genetic Linkage; glycosidases; immunoconjugate; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; N-glycoside hydrolases; nucleotide sequences; plant proteins; Plant Proteins - biosynthesis; Plant Proteins - genetics; Plants - genetics; Polysaccharide-Lyases - genetics; Promoter Regions, Genetic; Protein synthesis; protein synthesis inhibitors; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Ribosome Inactivating Proteins, Type 1; ribosomes; ricin; rna glycosidase; Sapindaceae; seeds; Sequence Homology, Amino Acid; structural genes; Transcription, Genetic; trichosanthin; aa; A; immunoconjugate; ricin; nt; RLA; G; n-Gel; trichosanthin; Gel; p; re; Protein synthesis; petB; RIP; oligo; PCR; araB; Complementary DNA; Nucleotide sequence; Dicotyledones; Angiospermae; Euphorbiaceae; Homology; Ribosome inactivating protein; Spermatophyta; Recombinant protein; Gene expression
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(93)90097-M

A cDNA copy of the gel gene, encoding gelonin (Gel), has been cloned from the seeds of the Asian plant, Gelonium multiflorum. Gel is a type-I ribosome-inactivating protein which has been produced in Escherichia coli as a secreted protein under the transcriptional control of the Salmonella typhimurium araB promoter and linked to the pectate lyase (pelB) leader sequence from Erwinia carotovora. Recombinant, soluble Gel (re-Gel) can be recovered from the E. coli culture supernatant at a yield of greater than 1 mg/ml, and it inhibits protein synthesis in vitro to the same extent as the native protein isolated from plant seeds.