Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis

XIAO Zheng; LI Ji-yuan; FAN Zheng-qi; YIN Heng-fu

Volume 27 Issue 3

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Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis

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

Received Date: 2013-07-17

Abstract

In this study, the actin gene was cloned from the stems of Camellia lipoensis using RT-PCR and RACE methods. The full length of the actin gene, named ClActin1, was 1 631 bp (GenBank accession No.KF366912), which contains a 1 134 bp open reading frame (ORF) encoding 377 amino acid residues, contains a 5'-UTR with 90 bp and 3'-UTR with 407 bp. The putative protein molecular weight is 41.69 kD and its theoretical isoelectric point is 5.31. The sequence includes actin superfamily domain and the characteristic actin family signature sequences. Homologous alignment shows that it shares over 82% of nucleotide identities and over 97% of amino acid identities with actins from other plants in GenBank. The phylogenetic tree constructed on the basis of amino acid sequences suggests that the relationship of actins from C. lipoensis is most intimate with that from C. sinensis and Populus trichocarpa. The results from the analysis on tissue specific expression show that the endogenous actin gene expression level in the root, stem, leaf and seed of C. lipoensis is identical. It can be served as inner control to determine the relative expression amount of target genes from Camellia plants.
- Camellia lipoensis,
- actin gene,
- sequence analysis,
- cloning

References

[1]	陈颖,王刚,赵俊霞. 高等植物体内的肌动蛋白[J]. 生物学通报,2003(1):13-15
[2]	朱筱娟,曾宪录,宋朝霞,等. 细胞核内肌动蛋白及其功能研究进展[J]. 科学通报,2004,49(11): 1031-1035
[3]	Thellin O, Zorzi W, Lakaye B, et al. Housekeeping genes as internal standards: use and limits[J]. Journal of Biotechnology, 1999, 75(2): 291-295
[4]	阎隆飞,石德权. 高等植物中的收缩蛋白[J]. 生物化学与生物物理学报, 1963, 3(4): 491-496
[5]	郭景康,陈青云,戢茜,等. 拟南芥、水稻和杨树ACTIN家族全基因组分析[J]. 上海大学学报: 自然科学版,2009,15(4): 426-431
[6]	Li X B, Fan X P, Wang X L, et al. The cotton ACTIN1 gene is functionally expressed in fibers and participates in fiber elongation[J]. The Plant Cell Online, 2005, 17(3): 859-875
[7]	陈鹏飞,刘雪梅,宋福南,等. 白桦肌动蛋白 (Actin) 基因全长 cDNA 克隆与序列分析[J]. 植物研究,2009,29(3): 339-345
[8]	马雄风,喻春明,唐守伟,等. 苎麻 Actin1 基因克隆及其在韧皮部纤维不同发育阶段的表达[J]. 作物学报,2010, 36(1): 101-108
[9]	王芳. 甘草肌动蛋白基因 GuActin2 的克隆和表达分析[J]. 植物生理学通讯, 2009,45(10): 995-1000
[10]	王舟,宗俊勤,宣继萍,等. 结缕草肌动蛋白基因全长cDNA的克隆及序列分析[J]. 草业学报,2010,19(6): 154-163
[11]	孔卫青,杨金宏. 桑树肌动蛋白actin基因全长序列的克隆与分析[J]. 广西植物, 2012,32(3): 362-366
[12]	阮桢媛,陈晓鸣,杨子祥. 角倍总RNA提取方法建立及ACTIN基因片段克隆[J]. 林业科学研究, 2012,25(5): 551-557
[13]	Wasteneys G O,Galway M E. Remodeling the cytoskeleton for growth and form: an overview with some new views[J]. Annual Review of Plant Biology, 2003, 54(1): 691-722
[14]	张宏达, 任善湘. 中国植物志: 第 49卷第3分册山茶科(一) 山茶亚科[M]. 北京: 科学出版社, 1998
[15]	Gasteiger E, Hoogland C, Gattiker A, et al. Protein identification and analysis tools on the ExPASy server[M]// Walker J M. The proteomics protocols handbook. Totowa,NJ: Humana Press Inc, 2005: 571-607
[16]	Bendtsen J D, Nielsen H, von Heijne G, et al. Improved prediction of signal peptides: SignalP 3.0[J]. Journal of Molecular Biology, 2004, 340(4): 783-795
[17]	Guermeur Y, Pollastri G, Elisseeff A, et al. Combining protein secondary structure prediction models with ensemble methods of optimal complexity[J]. Neurocomputing, 2004, 56: 305-327
[18]	Tamura K, Dudley J, Nei M, et al. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution, 2007, 24(8): 1596-1599
[19]	陈颖,赵俊霞,郝丽梅. 花粉肌动蛋白研究进展[J]. 生物学杂志,2002,(3): 5-8
[20]	Hightower R C, Meagher R B. The molecular evolution of actin[J]. Genetics, 1986, 114(1): 315-332
[21]	Perler F, Efstratiadis A, Lomedico P, et al. The evolution of genes: the chicken preproinsulin gene[J]. Cell, 1980, 20(2): 555-566
[22]	Fletcher L D, McDowell J M, Tidwell R R, et al. Structure, expression and phylogenetic analysis of the gene encoding actin1 in Pneumocystis carinii[J]. Genetics, 1994, 137(3): 743-750
[23]	Firtel R A. Multigene families encoding actin and tubulin[J]. Cell, 1981, 24(1): 6-7
[24]	Kandasamy M K, Burgos-Rivera B, McKinney E C, et al. Class-specific interaction of profilin and ADF isovariants with actin in the regulation of plant development[J]. The Plant Cell Online, 2007,19(10): 3111-3126
[25]	张少斌,刘国琴. 植物肌动蛋白异型体研究进展[J]. 植物学通报,2006(3): 242-248

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis

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

Received Date: 2013-07-17

Abstract: In this study, the actin gene was cloned from the stems of Camellia lipoensis using RT-PCR and RACE methods. The full length of the actin gene, named ClActin1, was 1 631 bp (GenBank accession No.KF366912), which contains a 1 134 bp open reading frame (ORF) encoding 377 amino acid residues, contains a 5'-UTR with 90 bp and 3'-UTR with 407 bp. The putative protein molecular weight is 41.69 kD and its theoretical isoelectric point is 5.31. The sequence includes actin superfamily domain and the characteristic actin family signature sequences. Homologous alignment shows that it shares over 82% of nucleotide identities and over 97% of amino acid identities with actins from other plants in GenBank. The phylogenetic tree constructed on the basis of amino acid sequences suggests that the relationship of actins from C. lipoensis is most intimate with that from C. sinensis and Populus trichocarpa. The results from the analysis on tissue specific expression show that the endogenous actin gene expression level in the root, stem, leaf and seed of C. lipoensis is identical. It can be served as inner control to determine the relative expression amount of target genes from Camellia plants.

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Reference (25)

[1]	陈颖,王刚,赵俊霞. 高等植物体内的肌动蛋白[J]. 生物学通报,2003(1):13-15
[2]	朱筱娟,曾宪录,宋朝霞,等. 细胞核内肌动蛋白及其功能研究进展[J]. 科学通报,2004,49(11): 1031-1035
[3]	Thellin O, Zorzi W, Lakaye B, et al. Housekeeping genes as internal standards: use and limits[J]. Journal of Biotechnology, 1999, 75(2): 291-295
[4]	阎隆飞,石德权. 高等植物中的收缩蛋白[J]. 生物化学与生物物理学报, 1963, 3(4): 491-496
[5]	郭景康,陈青云,戢茜,等. 拟南芥、水稻和杨树ACTIN家族全基因组分析[J]. 上海大学学报: 自然科学版,2009,15(4): 426-431
[6]	Li X B, Fan X P, Wang X L, et al. The cotton ACTIN1 gene is functionally expressed in fibers and participates in fiber elongation[J]. The Plant Cell Online, 2005, 17(3): 859-875
[7]	陈鹏飞,刘雪梅,宋福南,等. 白桦肌动蛋白 (Actin) 基因全长 cDNA 克隆与序列分析[J]. 植物研究,2009,29(3): 339-345
[8]	马雄风,喻春明,唐守伟,等. 苎麻 Actin1 基因克隆及其在韧皮部纤维不同发育阶段的表达[J]. 作物学报,2010, 36(1): 101-108
[9]	王芳. 甘草肌动蛋白基因 GuActin2 的克隆和表达分析[J]. 植物生理学通讯, 2009,45(10): 995-1000
[10]	王舟,宗俊勤,宣继萍,等. 结缕草肌动蛋白基因全长cDNA的克隆及序列分析[J]. 草业学报,2010,19(6): 154-163
[11]	孔卫青,杨金宏. 桑树肌动蛋白actin基因全长序列的克隆与分析[J]. 广西植物, 2012,32(3): 362-366
[12]	阮桢媛,陈晓鸣,杨子祥. 角倍总RNA提取方法建立及ACTIN基因片段克隆[J]. 林业科学研究, 2012,25(5): 551-557
[13]	Wasteneys G O,Galway M E. Remodeling the cytoskeleton for growth and form: an overview with some new views[J]. Annual Review of Plant Biology, 2003, 54(1): 691-722
[14]	张宏达, 任善湘. 中国植物志: 第 49卷第3分册山茶科(一) 山茶亚科[M]. 北京: 科学出版社, 1998
[15]	Gasteiger E, Hoogland C, Gattiker A, et al. Protein identification and analysis tools on the ExPASy server[M]// Walker J M. The proteomics protocols handbook. Totowa,NJ: Humana Press Inc, 2005: 571-607
[16]	Bendtsen J D, Nielsen H, von Heijne G, et al. Improved prediction of signal peptides: SignalP 3.0[J]. Journal of Molecular Biology, 2004, 340(4): 783-795
[17]	Guermeur Y, Pollastri G, Elisseeff A, et al. Combining protein secondary structure prediction models with ensemble methods of optimal complexity[J]. Neurocomputing, 2004, 56: 305-327
[18]	Tamura K, Dudley J, Nei M, et al. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution, 2007, 24(8): 1596-1599
[19]	陈颖,赵俊霞,郝丽梅. 花粉肌动蛋白研究进展[J]. 生物学杂志,2002,(3): 5-8
[20]	Hightower R C, Meagher R B. The molecular evolution of actin[J]. Genetics, 1986, 114(1): 315-332
[21]	Perler F, Efstratiadis A, Lomedico P, et al. The evolution of genes: the chicken preproinsulin gene[J]. Cell, 1980, 20(2): 555-566
[22]	Fletcher L D, McDowell J M, Tidwell R R, et al. Structure, expression and phylogenetic analysis of the gene encoding actin1 in Pneumocystis carinii[J]. Genetics, 1994, 137(3): 743-750
[23]	Firtel R A. Multigene families encoding actin and tubulin[J]. Cell, 1981, 24(1): 6-7
[24]	Kandasamy M K, Burgos-Rivera B, McKinney E C, et al. Class-specific interaction of profilin and ADF isovariants with actin in the regulation of plant development[J]. The Plant Cell Online, 2007,19(10): 3111-3126
[25]	张少斌,刘国琴. 植物肌动蛋白异型体研究进展[J]. 植物学通报,2006(3): 242-248

Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis

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通讯作者: 陈斌, bchen63@163.com

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Cloning and Expression Analysis of Full Length cDNA of Actin Gene from Camellia lipoensis

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