铁皮石斛促分裂原活化蛋白激酶基因dompk4的分离和差异表达分析.pdfVIP

1076 药学学报 Acta Pharmaceutica Sinica 20 14, 49 (7): 1076 1083 Isolation and differential expression of a novel MAP kinase gene DoMPK4 in Dendrobium offic inale 1, 2 1 1 2 2* ZHANG Gang , LI Yi-min , HU Ben-xiang , ZHANG Da-wei , GUO Shun-xing (1. College of Pharmacy, Shaanxi Provincial Key Laboratory f or Chinese Medicine Basis New Drugs Research, Shaanxi University of Chinese Medicine, Xi an 712046, China; 2. Institute of Medicinal Plant Development, Chinese A cademy of Medica l Sciences and Peking Union Medical College, Beij ing 100193, China ) Abstract : Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No . JX297597), is identified from a rare endangered medicinal orchid species D. offic inale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158 170), a MAP kinase site (71 174), and eight conserved motifs. DoMPK4 had a transmembrane (2 14 232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9% 80.6%) with MAPK proteins from various plants. Phylogenetic anal