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Molecular cloning and characterization of a mannose-binding lectin gene from Crinum asiaticum

文献类型: 外文期刊

作者: Chai, YR 1 ; Pang, YZ 1 ; Liao, ZH 1 ; Zhang, L 1 ; Sun, XF 1 ; Lu, YQ 1 ; Wang, SG 1 ; Tang, KX 1 ;

作者机构: 1.Shanghai Jiao Tong Univ, Fudan SJTU Nottingham Plant Biotechnol R&D Ctr, Plant Biotechnol Res Ctr, Sch Agr & Biol, Shanghai 200030, Peoples R China; SW Agr Univ, Sch Agron & Life Sci, Chongqing 400716, Peoples R China; Fudan Univ, SJTU Nottingham Plant Biotechnol R&D Ctr, State Key Lab Genet Engn, Sch Life Sci,Morgan Tan Int Ctr Life Sci, Shanghai 200433, Peoples R China; Sichuan Acad Agr Sci, Ind Crop Inst, Sichuan 641400, Peoples R China

关键词: Cdna cloning;Crinum asiaticum var. sinicum;Mannose-binding lectin;Transgenic plants;Antifungal protein;Brown planthopper;Glasshouse;trials;Gastrodia-elata;Resistance;Gna;Expression;Foods

期刊名称:JOURNAL OF PLANT PHYSIOLOGY ( 影响因子:3.549; 五年影响因子:4.164 )

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收录情况: SCI

摘要: Full-length c;DNA of a mannose-binding lectin or agglutinin gene was cloned from a traditional Chinese medicinal herb Crinum asiaticum var. sinicum through RACE-PCR cloning. The full-length cIDNA of C. asiaticum agglutinin (caa) was 820 bp and contained a 528 bp open reading frame encoding a lectin precursor (preproprotein) of 175 amino acid residues with a 22 aa signal peptide. The coding region of the caa gene was high in G/C content. The first 20 bp of the 5'UTR had a dC content of 50 %, which was a typical feature of the leader sequence. By cutting away the signal peptide, the CAA proprotein was 15.79 kDa with a pl of 9.27 and contained 3 mannose-binding sites (QDNY). Random coil and extended strand constituted interlaced domination of the main part of the secondary structure. B-lectin conserved domain existed within N-24 to G(130). Predicted three-dimensional structure of CAA proprotein was very similar to that of GNA (Galanthus nivalis agglutinin). It is significant that besides certain homologies to known monocot mannose-binding lectins from Amaryllidaceae, Orchidaceae, Alliaceae and Liliaceae, caa also showed high similarity to gastrodianin type antifungal proteins. No intron was detected within the region of genomic sequence corresponding to the caa full-length c;DNA. Southern blot analysis indicated that the caa gene belonged to a low-copy gene family. Northern blot analysis demonstrated that caa mRNA was constitutively expressed in all the tested tissue types including the root, bulb, leaf, rachise, flower and fruit tissues.

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