全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

绿竹BoGPIAP基因克隆与异位表达研究

Keywords: 绿竹 糖基化磷脂酰肌醇锚定蛋白基因 亚细胞定位 异位表达 纤维细胞

Full-Text   Cite this paper   Add to My Lib

Abstract:

糖基化磷脂酰肌醇锚定蛋白(GPIAP)因其结构和功能的多样性,决定了它在各种生物学过程中都发挥着重要作用。采用同源克隆的方法从绿竹(Bambusa oldhamii)中获得一个GPIAP同源基因,命名为BoGPIAP,cDNA全长1 772 bp,其中包括1 356 bp的开放阅读框,编码一个451 aa的的蛋白。蛋白结构分析表明,该蛋白包含1个典型的GPIAP家族保守区域(47-211)和1个CCVS结构域,在N-端和C-端分别具有1个跨膜信号肽和1个GPI锚定信号肽,属于GPIAP家族。构建BoGPIAP与GFP融合的表达载体,在洋葱表皮细胞中瞬时表达,结果显示BoCOBL::GFP融合蛋白定位于细胞膜上,证明BoGPIAP基因编码的蛋白为膜蛋白。分别构建BoGPIAP的正义、反义表达载体并转化烟草(Nicotiana tabacum)。PCR检测结果表明,BoGPIAP已转入烟草。与野生型相比,转反义基因植株细弱,纤维细胞壁明显变薄;而转正义基因植株粗壮,纤维细胞壁明显变厚。表明BoGPIAP可能对竹子纤维细胞壁的发育具有调控作用

References

[1]  Xiang J J, Zhang G H, Qian Q, et al. SEMI-ROLLEDLEAF1 encodes a putative glycosylphosphatidylinositol-anchored protein and modulates rice leaf rolling by regulating the formation of bulliform cells[J]. Plant Physiol, 2012, 159(4):1488-1500.
[2]  Li Y, Qian Q, Zhou Y, et al.BRITTLE CULM1, which encodes a COBRA-Like protein, affects the mechanical properties of rice plants[J]. Plant Cell, 2003, 15(9):2020-2031.
[3]  Liao S C, Lin C S, Wang A Y, et al. Differential expression of genes encoding acid invertases in multiple shoots of bamboo in response to various phytohormones and environmental factors[J]. J Agric Food Chem. 2013, 61(18):4396-405.
[4]  Yeh S H, Lee B H, Liao S C, et al. Identification of genes differentially expressed during the growth of Bambusa oldhamii[J]. Plant Physiol Biochem. 2013, 63:217-226.
[5]  Chen C Y, Hsieh M H, Yang C C, et al. Analysis of the cellulose synthase genes associated with primary cell wall synthesis in Bambusa oldhamii[J]. Phytochemistry, 2010, 71(11-12):1270-1279.
[6]  朱龙飞,徐英武,林新春. 绿竹花发育相关基因BoAP3的克隆与分析[J]. 浙江农林大学学报,2013, 30(6):839-842.
[7]  Lin X C, Huang L C, Fang W, et al. Understanding bamboo flowering based on large scale analysis of expressed sequence tags[J]. Gen Mol Res, 2010, 9(2):1085-1093.
[8]  Gao Z M, Li X P, Li L B, et al. An effective method for total RNA isolation from bamboo[J]. Chinese For Sci Tech, 2006, 5(3):52-54.
[9]  Kyte J, Doolittle R F. A simple method for displaying the hydropathic character of a protein[J]. J Mol Biol, 1982, 157, 105-132.
[10]  Sigrist C J, Cerutti L, de Castro E, et al. PROSITE, a protein domain database for functional characterization and annotation[J]. Nucleic Acids Res, 2010,38(Database issue):D161-6.
[11]  Geourjon C, Deléage G. SOPMA:Significant improvement in protein secondary structure prediction by consensus prediction from multiple alignments[J]. Comput Appl Biosci, 1995, 11(6), 681-684.
[12]  Hofmann K, Stoffel W. TMbase-A database of membrane spanning proteins segments[J]. Biol Chem, 1993, Hoppe-Seyler 374, 166.
[13]  Petersen T N, Brunak S, von Heijne G, et al. SignalP 4.0:discriminating signal peptides from transmembrane regions[J]. Nat Methods, 2011, 8(10):785-786.
[14]  Gao Z M, Zheng B, Wang W Y, et al. Cloning and functional characterization of a GNA-like lectin from Chinese Narcissus (Narcissus tazetta var. Chinensis Roem)[J]. Physiol Plantarum, 2011,142(2):193-204.
[15]  徐刚标,卢孟柱,陆燕. ARF基因导人烟草的遗传转化研究[J]. 中南林业科技大学学报,2007, 27(5):6-12.
[16]  曹双平.植物单根纤维拉伸性能测试与评价[D]. 中国林业科学研究院硕士论文,2010.
[17]  Eisenhaber B, Bork P, Eisenhaber F. Sequence properties of GPI-anchored proteins near the omega-site:constraints for the polypeptide binding site of the putative transamidase[J]. Protein Eng, 1998, 11(12):1155-1161.
[18]  Eisenhaber B, Bork P, Eisenhaber F. Prediction of potential GPI-modification sites in proprotein sequences[J]. J Mol Biol, 1999, 292(3):741-758.
[19]  Udenfriend S, Kodukula K. How glycosylphosphatidylinositol-anchored membrane proteins are made[J]. Annu Rev Biochem, 1995, 64:563-591.
[20]  刘兵. 水稻GPI锚定蛋白基因OsGPA1的生物学功能研究[D]. 中山大学博士学位论文,2009.
[21]  Brady S M, Song S, Dhugga K S, et al. Combining expression and comparative evolutionary analysis. The COBRA gene family[J]. Plant Physiol, 2007, 143(1):172-187.
[22]  Sedbrook J C, Carroll K L, Hung K F, et al. The ArabidopsisSKU5 gene encodes an extracellular glycosyl phosphatidylinositol-anchored glycoprotien involved in directional root growth[J]. Plant Cell, 2002, 14(7):1635-1648.
[23]  李雪平,高志民,彭镇华,等. 绿竹咖啡酸-O-甲基转移酶基因(COMT)的克隆及相关分析[J].林业科学研究,2007, 20(5):722-725.
[24]  金顺玉,卢孟柱,高健. 毛竹木质素合成相关基因C4H的克隆及组织表达分析[J]. 林业科学研究,2010, 23(3):319-325.
[25]  Gao Z M, Wang X C, Peng Z H, et al. Characterization and primary functional analysis of phenylalanine ammonia-lyase gene from Phyllostachys edulis[J]. Plant Cell Rep, 2012, 31(7):1345-1356.
[26]  Hsieh L S, Hsieh Y L, Yeh C S, et al. Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii[J]. Mol Biol Rep, 2011, 38(1):283-290.
[27]  Hsieh L S, Yeh C S, Pan H C, et al. Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris[J]. Protein Expr Purif, 2010, 71(2):224-30.
[28]  杜亮亮,鲁专,金爱武. 雷竹纤维素合成酶基因cDNA克隆与表达分析[J]. 江西农业大学学报,2010, 32(3):535-540.
[29]  Chiu W B, Lin C H, Chang C J, et al. Molecular characterization and expression of four cDNAs encoding sucrose synthase from green bamboo Bambusa oldhamii[J]. New Phytol, 2006, 170(1):53-63.
[30]  高志民,杨学文,彭镇华,等.绿竹BoSUT2基因的克隆、分析与亚细胞定位研究[J]. 林业科学,2010, 46(2):45-49.
[31]  Roudier F, Fernandez A G, Fujita M, et al. COBRA, an Arabidopsis extracellular glycosyl-phosphatidyl inositol-anchored protein, specifically controls highly anisotropic expansion through its involvement in cellulose microfibril orientation[J]. Plant Cell, 2005, 17(6):1749-1763.
[32]  Wasteneys G O, Fujita M. Establishing and maintaining axial growth:wall mechanical properties and the cytoskeleton[J]. J Plant Res, 2006, 119(1):5-10.
[33]  Sindhu A, Langewisch T, Olek A, et al. Maize Brittle stalk2 encodes a COBRA-like protein expressed in early organ development but required for tissue flexibility at maturity[J]. Plant Physiol, 2007, 145(4):1444-1459.
[34]  Hsieh L S, Yeh C S, Pan H C, et al. Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris[J]. Protein Expr Purif, 2010, 71(2):224-30.
[35]  杨学文,彭镇华. 一个毛竹细胞色素P450基因的克隆与表达研究[J]. 安徽农业大学学报,2010, 37(1):116-121.
[36]  Shi H, Kim Y, Guo Y, et al. The ArabidopsisSOS5 locus encodes a putative cell surface adhesion protein and is required for normal cell expansion[J]. Plant Cell, 2003, 15(1):19-32.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133