The sequence of 5' cDNA ends of Betaine-aldehyde Dehydrogenase(BADH)was cloned from Puccinellia chinampoensis by improved RT-PCR and 5' RACE methods.The full length of cDNA was 1 781 bp by overlapping sequence and contained an intact open reading frame(ORF)encoding a putative protein of 506 amino acids.The deduced amino acid sequence contained the conserved motif of aldehyde dehydrogenase which was VT/SLELGGKSP and after it there was the Cys at the 29th site,associated with aldehyde dehydrogenase function.Multiple sequence alignment and phylogenetic analysis showed that the polypeptide shared high homology with Hordeum brevisubulatum BBD1.Then its plant expression vector named PBI121-BADH was constructed.
WANG Lei
,
ZHONG Ming
,
GUO Zhi-fu
,
ZHANG Li
,
ZHANG Jia
,
ZHAO Li
,
LI Hao-ge
. Cloning of 5' cDNA Ends of Betaine Aldehyde Dehydrogenase from Puccinellia chinampoensis and Construction of Plant Expression Vector[J]. Acta Agriculturae Boreali-Sinica, 2010
, 25(2)
: 30
-34
.
DOI: 10.7668/hbnxb.2010.02.006
[1] 梁峥,骆爱玲.甜菜碱和甜菜碱合成酶[J].植物生理学通讯,1995,31(1):1-8.
[2] Sakamoto A,Murata N.Genetic engineering of glycinebetaine synthesis in plants:current status and implications for enhancement of stress tolerance[J].J Exp Bot,2000,51(342):81-88.
[3] Weretilnyk E A,Hanson A D.Molecular cloning of a plant betaine aldehyde dehydrogenase,an enzyme implicated in adaptation to salinity and drought[J].Proc Natl Acad Sci USA,1990,87(7):2745-2749.
[4] McCue K F,Hanson A D.Salt-inducible betaine aldehyde dehydrogenase from sugar beet:cDNA cloning and expression[J].Plant Mol Biol,1992,18(1):1-11.
[5] 白杰英,陆一鸣,庞晓斌,等.盐胁迫短芒大麦甜菜碱醛脱氢酶基因片段的克隆与序列分析[J].内蒙古民族大学学报:自然科学版,2003,18(1):38-41.
[6] Li Q L,Gao X R,Yu X H,et al.Molecular cloning and characterization of betaine aldehyde dehydrogenase gene from Suaeda liaotungensis and its use in improved tolerance to salinity in transgenic tobacco[J].Biotechnol Lett,2003,25(17):1431-1436.
[7] 何晓兰,侯喜林,吴纪中,等.三角叶滨藜甜菜碱醛脱氢酶(BADH)基因的克隆及序列分析[J].南京农业大学学报,2004,27(1):15-19.
[8] Oishi H,Ebina M.Isolation of cDNA and enzymatic properties of betaine aldehyde dehydrogenase from Zoysia tenuifolia[J].J Plant Physiol,2005,162:1077-1086.
[9] 徐恒刚.中国盐生植被及盐渍化生态治理[M].北京:中国农业科学技术出版社,2003:244-245.
[10] 钟鸣,张佳,郭志富,等.朝鲜碱茅BADH基因3'端及5'端部分序列的扩增[J].华北农学报,2009,24(3):46-50.
[11] 刘振林,戴思兰.植物甜菜碱醛脱氢酶基因研究进展[J].西北农林科技大学学报:自然科学版,2004,32(3):104-112.
[12] McCue K F,Hanson A D.Salt-inducible betaine aldehyde dehydrogenase from sugar beet:cDNA cloning and expression[J].Plant Mo lecular Biology,1992,18:1-11.
[13] Nakamura T,Nomura M,Mori H,et al.An isozyme of betaine aldehyde dehydrogenase in barley[J].Plant Cell Physiol,2001,42(10):1088-1092.
[14] 杨峥,钟鸣,郭志富,等.盐胁迫下朝鲜碱茅的甜菜碱醛脱氢酶活性变化及其基因保守区的克隆[J].植物生理学通讯,2007,43(3):430-434.
[15] Papp A C,Snyder P J,Sedra M,et al.Strategies for amplification of trinucleotide repeates:optimization of fragile X and androgen receptor PCR[J].Mol Diagn,1996,1(1):59-64.
[16] Henke W,Herdel K.Betion improves the PCR amplification of GC-rich DNA sequences[J].Nucleic Acids Rcs,1997,25(19);3957-3958.