通过叶绿体基因工程在烟草中合成中长链羟基脂肪酸聚酯

被引:3
作者
王玉华
张秀海
吴忠义
杨清
陈国强
吴琼
黄丛林
机构
[1] 南京农业大学生命科学学院
[2] 北京市农林科学院北京农业生物技术研究中心
[3] 清华大学生物科学与技术系
[4] 北京市农林科学院北京农业生物技术研究中心 南京 北京市农林科学院北京农业生物技术研究中心
关键词
中长链羟基脂肪酸聚酯; 叶绿体基因工程; 转基因烟草; 植物生物反应器;
D O I
暂无
中图分类号
S572 [烟草(菸草)];
学科分类号
摘要
中长链羟基脂肪酸聚酯(medium-chain-length-PHAs,mcl-PHAs)属于微生物聚酯.PHA合酶(PHAsynthase,由phaC基因编码)和3-羟酰基-酰基转移蛋白-辅酶A转移酶(3-hydroxyacyl-acylcarrierprotein-coenzymeAtransferase,由phaG基因编码)是mcl-PHAs生物合成途径中的关键酶.以aadA(氨基糖苷-3′-磷酸转移酶)基因做筛选标记,分别构建了含phaC2基因的单价叶绿体转化表达载体pTC2以及同时嵌合phaC和phaG基因的双价叶绿体转化表达载体pTGC.通过PDS1000/He基因枪转化法转化烟草,获得具有壮观霉素抗性的叶绿体型转基因烟草植株,PCR和SouthernBlot结果表明,外源基因确已整合进烟草叶绿体基因组中且基本达到同质化.气相色谱法检测转基因株系中的mcl-PHAs含量最高达4.8mg/g干重,合成的PHAs单体主要成分为3-羟基辛酸(3-hydroxyoctanoate,3-HO)和3-羟基癸酸(3-dydroxydecanoate,3-HD).透射电子显微镜观察到转基因烟草叶片叶绿体中确有PHAs颗粒累积.
引用
收藏
页码:979 / 986
页数:8
相关论文
共 10 条
  • [1] Expression of poly-3-(R)-hydroxyalkanoate (PHA) polymerase and acyl-CoA-transacylase in plastids of transgenic potato leads to the synthesis of a hydrophobic polymer, presumably medium-chain-length PHAs
    Romano, A
    van der Plas, LHW
    Witholt, B
    Eggink, G
    Mooibroek, H
    [J]. PLANTA, 2005, 220 (03) : 455 - 464
  • [2] Transgene organisation in potato after particle bombardment-mediated (co-)transformation using plasmids and gene cassettes
    Romano, A
    Raemakers, K
    Bernardi, J
    Visser, R
    Mooibroek, H
    [J]. TRANSGENIC RESEARCH, 2003, 12 (04) : 461 - 473
  • [3] Polyester synthesis in transplastomic tobacco (Nicotiana tabacum L.):: significant contents of polyhydroxybutyrate are associated with growth reduction
    Lössl, A
    Eibl, C
    Harloff, HJ
    Jung, C
    Koop, HU
    [J]. PLANT CELL REPORTS, 2003, 21 (09) : 891 - 899
  • [4] Expression of bacterial poly (3-hydroxybutyrate) synthesis genes in hairy roots of sugar beet (Beta vulgaris L.)
    Menzel, G
    Harloff, HJ
    Jung, C
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2003, 60 (05) : 571 - 576
  • [5] Poly(β-hydroxybutyrate) production in oilseed leukoplasts of Brassica napus
    Houmiel, KL
    Slater, S
    Broyles, D
    Casagrande, L
    Colburn, S
    Gonzalez, K
    Mitsky, TA
    Reiser, SE
    Shah, D
    Taylor, NB
    Tran, M
    Valentin, HE
    Gruys, KJ
    [J]. PLANTA, 1999, 209 (04) : 547 - 550
  • [6] Updated gene map of tobacco chloroplast DNA
    Wakasugi, T
    Sugita, M
    Tsudzuki, T
    Sugiura, M
    [J]. PLANT MOLECULAR BIOLOGY REPORTER, 1998, 16 (03) : 231 - 241
  • [7] The complete sequence of the rice ( Oryza sativa ) chloroplast genome: Intermolecular recombination between distinct tRNA genes accounts for a major plastid DNA inversion during the evolution of the cereals.[J].Junzou Hiratsuka;Hiroaki Shimada;Robert Whittier;Takashi Ishibashi;Masahiro Sakamoto;Masao Mori;Chihiro Kondo;Yasuko Honji;Chong-Rong Sun;Bing-Yuan Meng;Yu-Qing Li;Akira Kanno;Yoko Nishizawa;Atsushi Hirai;Kazuo Shinozaki;Masahiro Sugiura.MGG Molecular & General Genetics.1989, 2
  • [8] 实用生物电子显微术.[M].林钧安等主编;.辽宁科学技术出版社.1989,
  • [9] 通过叶绿体基因工程表达聚3-羟基丁酸酯合成相关基因
    张景昱
    赵华燕
    朱生伟
    宋艳茹
    苏宁
    张中林
    [J]. 科学通报, 2002, (11) : 845 - 848
  • [10] 高等植物叶绿体DNA提纯方法的改进
    龚小松
    阎隆飞
    不详
    [J]. 科学通报 , 1991, (06) : 467 - 469