Cloning and Sequence Analysis of Full-length cDNA of
pepc Gene from Jatropha curcas

FAN Zheng-qi; LI Ji-yuan; TIAN Min; LI Xin-lei; CHEN Dong-liang; LU Meng-zhu

Applying RT-PCR and RACE method, 3 142 bp pepc gene was isolated from Jatropha curcas. The sequence analysis showes that the pepc gene including an open reading frame is 2 898 bp in length, and encodes a protein of 965 amino acid. Its amino acid sequence shares 94.94%, 92.46%, 90.60%, 90.50%, 90.50%, 88.33%, 84.61%, 82.44% identity with those of pepc from Ricinus communis, Gossypium hirsutum, Citrus sinensis, Glycine max, Arachis hypogaea, Nicotiana tabacum, Brassica napus, Arabidopsis thaliana, respectively. The pepc gene has been accepted by GenBank, and the accession number of gene sequence is EU069413. The deduced protein molecular weight of pepc is 110.6 kD, and belong to pepc1 in gene family and C3 form. Its stability, secondary structure, hydrophobicity profile were analysed, and the functional sites and characterized domains were confirmed.

Cloning and Sequence Analysis of Full-length cDNA of pepc Gene from Jatropha curcas

1. Research Institute of Subtropical Forestry, Chinese Academy Forestry, Fuyang 311400, Zhejiang, China

2. Research Institute of Forestry, Chinese Academy Forestry , Beijing 100091, China)

Abstract: Applying RT-PCR and RACE method, 3 142 bp pepc gene was isolated from Jatropha curcas. The sequence analysis showes that the pepc gene including an open reading frame is 2 898 bp in length, and encodes a protein of 965 amino acid. Its amino acid sequence shares 94.94%, 92.46%, 90.60%, 90.50%, 90.50%, 88.33%, 84.61%, 82.44% identity with those of pepc from Ricinus communis, Gossypium hirsutum, Citrus sinensis, Glycine max, Arachis hypogaea, Nicotiana tabacum, Brassica napus, Arabidopsis thaliana, respectively. The pepc gene has been accepted by GenBank, and the accession number of gene sequence is EU069413. The deduced protein molecular weight of pepc is 110.6 kD, and belong to pepc1 in gene family and C3 form. Its stability, secondary structure, hydrophobicity profile were analysed, and the functional sites and characterized domains were confirmed.

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Reference (21)

[1]	Gregory A L, Hurley B A, Tran H T, et al. In vivo regulatory phosphorylation of the phosphoenolpyruvate carboxylase AtPPC1 in phosphate-starved Arabidopsis thaliana[J]. Biochem J, 2009, 420(1): 57-65
[2]	Takahashi-Terada A, Kotera M, Ohshima K, et al. Maize phosphoenolpyruvate carboxylase: mutations at the putative binding site for glucose 6-phosphate caused desensitization and abolished responsiveness to regulatory phosphorylation[J]. J Biol Chem, 2005, 280(12): 11798-11806
[3]	Cousins A B, Adam N R, Wall G W, et al.Development of C₄ photosynthesis in sorghum leaves grown under free-air CO₂ enrichment (FACE)[J]. Journal of Experimental Botany, 2003, 54(389): 1969-1975
[4]	Gennidakis S, Rao S, Greenham K, et al.Bacterial- and plant-type phosphoenolpyruvate carboxylase polypeptides interact in the hetero-oligomeric Class-2 PEPC complex of developing castor oil seeds[J]. Plant J, 2007, 52(5): 839-849
[5]	Alvarez R, Castillo J M, Mateos-Naranjo E,et al.Ecotypic variations in phosphoenolpyruvate carboxylase activity of the cordgrass Spartina densiflora throughout its latitudinal distribution range[J]. Plant Biology, 2009, 12(1): 154-160
[6]	Wang X, Gowik U, Tang H, et al. Comparative genomic analysis of C₄ photosynthetic pathway evolution in grasses[J]. Genome Biol 2009, 10(6):68
[7]	张桂芳, 赵明, 丁在松, 等. 稗草磷酸烯醇式丙酮酸羧化酶(PEPCase)基因的克隆与分析[J]. 作物学报, 2005, 31(10): 1365-1369
[8]	Sugimoto T, Tanaka K. Phosphoenolpyruvate carboxylase level in soybean seed highly correlates to its contents of protein and lipid[J]. Agric Biol Chem, 1989, 53(3): 885-887
[9]	陈锦清, 黄锐之, 郎春秀, 等. 油菜PEP基因的克隆及PEP反义基因的构建[J]. 浙江大学学报:农业与生命科学版, 1999, 25(4):365-367
[10]	郎春秀, 胡张华, 刘智宏, 等. 油菜农杆菌转基因体系的建立及转PEP反义基因油菜的获得[J]. 浙江农业学报, 1999, 11(2):55-58
[11]	陈锦清, 郎春秀, 胡张华, 等. 反义PEP基因调控油菜籽粒蛋白质/油脂含量比率的研究[J]. 农业生物技术学报, 1999, 7(4):316-320
[12]	候李君, 施定基, 蔡泽富, 等. 蓝藻正反义pepcA基因导入对大肠杆菌中脂类合成的调控[J]. 中国生物工程杂志, 2008, 28(5):52-58
[13]	赵桂兰, 陈锦清, 尹爱萍, 等. 获得转反义PEP基因超高油大豆新材料[J]. 分子植物育种, 2005, 3(6):792-796
[14]	吴关庭, 郎春秀, 胡张华, 等. 应用反义PEP基因表达技术提高稻米脂肪含量[J]. 植物生理与分子生物学学报, 2006, 32(3):339-344
[15]	郝乃斌, 戈巧英, 杜维广. 大豆高光效育种光合生理研究进展[J]. 植物学通报, 1991, 8(2): 13-19
[16]	Westhoff P, Gowik U. Evolution of C₄ phosphoenolpyruvate carboxylase genes and proteins: a case study with the genus Flaveria[J]. Annals of Botany, 2004, 93: 13-23
[17]	张边江, 华春, 周峰, 等. 转PEPC+PPDK双基因水稻的光合特性[J]. 中国农业科学, 2008, 41(10): 3008-3014
[18]	丁在松, 赵明, 荆玉祥, 等. 玉米ppc基因过表达对转基因水稻光合速率的影响[J]. 作物学报, 2007,33(5): 717-722
[19]	张彬, 丁在松, 张桂芳, 等. 根癌农杆菌介导获得稗草Ecppc转基因小麦的研究[J]. 作物学报, 2007, 33(3): 356-362
[20]	陈绪清, 张晓东, 梁荣奇, 等. 玉米C₄型pepc基因的分子克隆及其在小麦的转基因研究[J]. 科学通报, 2004, 49(19): 1976-1982
[21]	王治涛. 麻疯树(Jatropha curcas L.)胚乳cDNA文库的构建及磷酸烯醇式丙酮酸羧化酶基因克隆的研究 . 成都:四川大学,2006

Cloning and Sequence Analysis of Full-length cDNA of pepc Gene from Jatropha curcas

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References

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