Cloning of XET gene from Anthocephalus chinensis and its plant expression vector construction

作     者：MA Sheng-jun ZHU Song-lin LI Wei OUYANG Kun-xi LI Na CHEN Xiao-yang 

作者机构：Key Laboratory for Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education National Engineering Laboratory for Forest Tree Breeding Beijing Forestry University Beijing 100083 P. R. China State-owned Forest Administration of Lvliangshan Mountain Linfen 041000 P. R. China Forestry Research Institute of Hebei Province Shijiazhuang 050061 P. R. China College of Forestry South China Agricultural University Guangzhou 200064 P. R. China 

出 版 物：《Forestry Studies in China》 (中国林学（英文版）)

年 卷 期：2010年第12卷第2期

页      面：79-84页

学科分类：0710[理学-生物学] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 0907[农学-林学] 07[理学] 08[工学] 0829[工学-林业工程] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 0713[理学-生态学] 

基　　金：supported by the National Natural Science Foundation of China (Grant No. 30901158) the Key Project of Chinese Ministry of Education (Grant No. 104243) 

主　　题：cDNA cloning sequence analysis AcXET gene plant expression vector 

摘      要：A full-length cDNA sequence of xyloglucan endotransglycosylase gene （XET）, abundantly expressed in the cambium of Anthocephalus chinensis was cloned by conserved PCR, rapid-amplification of cDNA ends and by chromosome walking. Analytical results of the DNA sequence show that a 912 bp complete open reading frame （ORF） encoded a 303-amino acid protein was in the 1205 bp full cDNA sequence. The deduced amino acid sequence of AcXET, which contained the conserved specific EIDFE catalytic site sequence to XETs was homologous to the other known XET proteins. In order to study the gene function of AcXET and obtain transgenic plants, a plant expression vector pBIAcXET was constructed by recombinating the AcXET fragment from the cloning vector pMD19AcXET and the binary vector pBI121 between the XbaI and SmaI sites. The fragment ofAcXET gene was inserted between the CaMV 35S promotor and the coding region of the GUS gene in pBI121. The identification results show that the plant expression binary vector pBIAcXET was constructed successfully. These results lay the foundation for studying the molecular mechanism ofAcXET gene during wood formation.