[1]宋丽雅,盖 燕,何聪芬.向大豆导入琉璃苣Δ6-脂肪酸脱氢酶基因的初步研究[J].大豆科学,2012,31(02):173-177.[doi:10.3969/j.issn.1000-9841.2012.02.003]
　SONG Li-ya,GE Yan,HE Cong-fen.A Preliminary Study on Transferring Δ6-Fatty Acid Desaturase Gene from Borago Officinalis into Soybean[J].Soybean Science,2012,31(02):173-177.[doi:10.3969/j.issn.1000-9841.2012.02.003]
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向大豆导入琉璃苣Δ6-脂肪酸脱氢酶基因的初步研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第31卷 期数: 2012年02期 页码: 173-177 栏目: 出版日期: 2012-04-25
Title:
A Preliminary Study on Transferring Δ6-Fatty Acid Desaturase Gene from Borago Officinalis into Soybean
文章编号:
1000-9841（2012）02-0173-05
作者:
宋丽雅; 盖 燕; 何聪芬
北京工商大学 北京市植物资源研究开发重点实验室，北京 100048
Author(s):
SONG Li-ya; GE Yan; HE Cong-fen
Beijing Technology and Business University,Beijing Key Laboratory of Plant Resources Research and Development，Beijing 100048,China
关键词:
γ-亚麻酸; 大豆; Δ6-脂肪酸脱氢酶基因; 农杆菌介导
Keywords:
γ-linolenic acid; Soybean; Δ6-fatty acid desaturase gene; Agrobacterium mediated
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2012.02.003
文献标志码:
A
摘要:
采用RT-PCR法扩增出中国境内生长的琉璃苣Δ6-脂肪酸脱氢酶基因，序列分析表明，该基因全长为1 359个核苷酸，与美国德克萨斯州的琉璃苣Δ6-脂肪酸脱氢酶基因同源性可达到99%。将克隆到的基因连接pRTL2质粒的35S启动子，构建重组载体pPTN-D6D，并通过农杆菌介导的子叶节转化系统，将该基因导入到垦农18和小粒豆2个大豆品种中。经PCR检测分析，初步证明琉璃苣Δ6-脂肪酸脱氢酶基因已导入并整合到大豆基因组中。
Abstract:
γ-linolenic acid is an essential polyunsaturated fatty acid in human metabolism.Δ6-fatty acid desaturase(D6D)is the key enzyme that can convert linoleic acid into γ-linolenic acid in vivo.In this study,the Δ6-fatty acid desaturase gene from Borago officinalis was amplified by RT-PCR.Sequence analysis showed that the full cDNA contained 1 359 nucleotides.The homology of Borago officinali ?D6D gene in China and that registered in Texas USA was 99%.Recombinant vector pPTN-D6D was constructed that containing the Δ6-fatty acid desaturase gene from Borago officinalis and 35S promoter from plasmid pRTL2.The gene was transferred into two soybean cultivars,Xiaolidou and Kennong18, with Agrobacterium mediated cotylendon node transformation system.The results of PCR indicated that the D6D gene had been introduced and integrated into soybean genome which laid a foundation for soybean quality improvement.

参考文献/References:

[1]卜云萍,王光风,陈竺.转基因高γ-亚麻酸大豆油抗肿瘤作用的研究[J].中国生物工程杂志，2005，25(7):92-97.(Bu Y P,Wang G F,Chen Z.Antitumor of the transgenic soybean oil[J].China Biotechnology,2005,25(7):92-97.)

[2]Murphy D J.Engineering oil production in rapeseed and other oil crops[J].Trends in Biotechnology,1996,14(6):206-213.

[3]Ohlrogge J B.Design of new plant products:engineering of fatty acid metabolism[J].Plant Physiology,1994,104:821-826.

[4]Giamarellos-Bourboulis E J,Grecka P,Dionyssiou-Asteriou A,et al.In vitro influence of polyunsaturated fatty acids on nosocomial Pscudomonas aerugi-nosa:a preliminary report[J].Inte Antimierob Agents,1995,6(1):47-50.

[5]武小霞,李文滨,张淑珍.我国大豆转基因研究进展[J].大豆科学，2005,24（2）:145-149.(Wu X X,Li W B,Zhang S Z.The research advance on soybean transgene in China[J].Soybean Science,2005,24（2）:145-149.)

[6]白义春,康相涛,孙桂荣,等.Δ6-脂肪酸脱氢酶研究进展[J].河南农业科学,2008(2):4-12.(Bai Y C,Kang X T,Sun G R,et al.Advance in Δ6-fatty acid desaturase research[J].Journal of Henan Agricultural Sciences,2008(2):4-12.)

[7]Khang N T,Jennen D G,Tholen E,et al.Association of the FADS2 gene with omega-6 and omega-3 PUFA concentration in the egg yolk of Japanese quail[J].Animal Biotechnology,2007,18(3):189-201.

[8]王广科,李明春,李晋川,等.高山被孢霉Δ6-脂肪酸脱氢酶基因转化大豆的研究[J].成都医学院学报,2010,5(2):93-96.(Wang G K,Li M C,Li J C,et al.The study on transformation of Δ6-fatty acid desaturase genes from Mortierella alpine?into soybean[J].Journal of Chengdu Medical College,2010,5(2):93-96.)

[9]张琦,李明春,蔡易,等.少根根霉Δ6-脂肪酸脱氢酶基因在转基因油菜中的表达[J].中国农业科学,2006,39(3):463-469.(Zhang Q,Li M C,Cai Y.et al.Expression of Rhizopus arrhizus?Δ6-fatty acid desaturase gene in transgenic rapeseed[J].Scientia Agricultura Sinica,2006,39(3):463-469.)

[10]李明春,刘莉,胡国武,等.深黄被孢霉Δ6-脂肪酸脱氢酶基因在转基因烟草中的表达[J].生物工程学报,2003,19(2):178-184.(Li C M,Liu L,Hu G W,et al.Expression of Mortierella isabellina?Δ6-fatty acid desaturase gene in γ-linolenic acid production in transgenic tobacco[J].Chinese Journal of Biotechnology,2003,19(2):178-184.)

[11]苗兴芬,徐文平,李灿东,等.东北地区大豆品种脂肪酸组成与含量分析[J].大豆科学,2011,30(1):529-531.(Miao X F,Xu W P,Li C D,et al.Analysis of the fatty acid composition of soybean varieties in northeast China[J].Soybean Science,2011,30(1):529-531.)

[12]李文滨,郑宇宏,韩英鹏.大豆种质资源脂肪酸组分含量及品质性状的相关性分析[J].大豆科学,2008,27(5):740-745.(Li W B,Zheng Y H,Han Y P.Analysis of fatty acid composition and other quality traits in soybean varieties developed in Heilongjiang province[J].Soybean Science,2008,27(5):740-745.)

[13]Messina M,Messina V L,Chan P.Soyfoods,hyperuricemia and gout:a review of the epidemiologic and clinical data[J].Asia Pacific Journal of Clinical Nutrition,2011,20(3):347-358.

[14]徐香玲,邹联沛.向大豆导入几丁质酶基因的初步研究[J].大豆科学,1999,18(2):101-108.(Xu X L,Zou L P.A preliminary study on transferring chitinase gene into soybeans.[J].Soybean Science,1999,18(2):101-108.)

[15]陈向明.用CTAB法提取植物DNA的技术改进[J].合肥教育学院学报,2000,17(4):14-16.(Chen X M.The technology improvement of extracting DNA from plants by CTAB[J].Journal of Hefei Institute of Education,2000,17(4):14-16.)

[16]Reddy A S,Nuccio M L,Rross L M,et al.Isolation of a Δ6-desaturase gene from the cyanobacterium Synechocystis?sp.strain PCC 6803 by gain-of-function expression in?Anabaena?sp.strain PCC 7120[J].Plant Molecular Biology,1993,22(2):293-300.

[17]刘睿杰.应用RACE技术克隆月见草Δ6-脂肪酸脱氢酶基因cDNA[D].呼和浩特：内蒙古大学，2007:1-35.(Liu R J.Cloning of Δ6 fatty acid desaturase gene cDNA from Oenothera biennis?using technology of race[D].Hohhot:Inner Mongolia University,2007:1-35.)

[18]王德培,李明春,魏东盛,等.雅致枝霉Δ6-脂肪酸脱氢酶基因的克隆及在酿酒酵母中的表达[J].微生物学报,2006,46(1):74-79.(Wang D P,Li M C,Wei D S,et al.Cloning and expression of Δ6-desaturase gene from Thamnidium elegans?in Saccharomyces cerevisiae[J].?Acta Microbiologica Sinica,2006,46(1):74-79.)

[19]Mauro A O,Pfeoffer T W,Collies G B.Inheritance of soybean susceptibility to Agrobacterium umefaciens?and its relationship to transformation[J].Crop Science,1995,35:1152-1156.

[20]王罡,王萍,蔺宇,等.大豆基因型对根癌农杆菌菌株敏感性的研究[J].遗传,2002,24(3):297-300.(Wang G,Wang P,Lin Y,et al.The studies of sensitivity of genotypes in soybean to lines of Agrobacterium tumefaciens[J].Hereditas,2002,24(3):297-300.)

[21]张书利,刘丽君,唐晓飞,等.利用农杆菌介导法将BrCS基因导入大豆的研究[J].大豆科学,2011,30(4):569-573.(Zhang S L,Liu L J,Tang X F.et al.Transformation of BrCS?gene into soybean by Agrobacterium-mediated method[J].Soybean Science,2011,30(4):569-573.)

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备注/Memo

基金项目：北京市教委科技发展计划项目（KM2009011008）；北京市自然科学基金项目（6112003）。

第一作者简介：宋丽雅（1974-），女，博士，讲师， 研究方向为酶工程。E-mail：songly@th.btbu.edu.cn。
通讯作者：何聪芬（1966-），女，博士，教授，研究方向为植物分子生物学。E-mail：hecf@th.btbu.edu.cn。

更新日期/Last Update: 2014-08-15