Isolation and Characterization of a Wound-inducible Ribonuclease fromNicotiana glutinosaLeaves

Molecular Cloning of cDNAs Encoding Ribonuclease-related Proteins inNicotiana glutinosaLeaves, as Induced in Response to Wounding or to TMV-infection

“…We earlier puriˆed a wound-inducible RNase NW from leaves of N. glutinosa to apparent homogeneity: 10) the former name``RNase M'' was changed to avoid confusion with a fungal RNase M from Aspergillus saito. In addition, a cDNA clone (NGR1 ) encoding RNase NW was ampliˆed and the nucleotide sequence was analyzed.…”

“…In addition, a cDNA clone (NGR1 ) encoding RNase NW was ampliˆed and the nucleotide sequence was analyzed. 10) Sequence comparison demonstrated that RNase NW is signiˆcantly related to S-like RNases, such as RNase NE from N. alata 11) and RNase LE 12) from cultured cells of tomatoes rather than to S-RNases. 10) This analysis also showed that RNase NW is synthesized as a polypeptide with 229 amino acids, including a putative secretion signal peptide at the N-terminus composed of 25 amino acids.…”

“…10) In this study, to acquire more information on RNases in N. glutinosa leaves, we ampliˆed two distinct full-length cDNAs, NGR2 and NGR3, encoding ribonucleaserelated proteins in the N. glutinosa leaves and analyzed expression patterns of transcripts in response to TMV-infection.…”

“…rapid ampliˆcation of cDNA ends (RACE) were done as previously described. 10) Cloning of 5? ends of cDNA were done using a 5?-Full RACE Core Set (Takara Shuzo) and ampliTaq Gold (Perkin-Elmer) according to the manufacturer's instructions.…”

“…Analysis of several independent 3? RACE products showed that transcripts from NGR2 and NGR3 genes The deduced amino acid sequences of NGR2 and NGR3 were aligned with those of RNase NW from N. glutinosa, 10) RNase NE from N. alata, 11) RNS1, RNS2, and RNS3 from A. thaliana, 17) RNases LE 12) and LX 18) from tomato (Lycopersicon esculantum ), and RNase MC1 from the bitter gourd (Momordica charantia) seeds. 20) The completely conserved residues are boxed.…”