Synthesis of medium-chain-length-polyhydroxyalkanoates in tobacco via chloroplast genetic engineering  

作　　者：WANGYuhua WUZhongyi ZHANGXiuhai CHENGuoqiang WUQiong HUANGConglin YANGQing 

机构地区：[1]BeijingAgro-BiotechnologyResearchCenter,BeijingAcademyofAgricultureandForestrySciences,Beijing100089,China//CollegeofLifeSciences,NanjingAgriculturalUniversity,Nanjing210095,China [2]BeijingAgro-BiotechnologyResearchCenter,BeijingAcademyofAgricultureandForestrySciences,Beijing100089,China [3]DepartmentofBiologicalSciencesandBiotechnology,TsinghuaUnversity,Beijing100084,China [4]CollegeofLifeSciences,NanjingAgriculturalUniversity,Nanjing210095,China

出　　处：《Chinese Science Bulletin》2005年第11期1113-1120,共8页

基　　金：the National 863 Program of China(Grant No.2002AA213051).

摘　　要：Medium-chain-length-polyhydroxyalkanoates (mcl-PHAs) belong to the group of microbial polyesters con- taining monomers ranging from 6 to 14 carbons in length. The key enzymes of their biosynthesis are PHA-polymerase (product of phaC gene) and 3-hydroxyacyl-acyl carrier pro- tein-CoA transferase (product of phaG gene). With aadA (aminoglycoside 3′-adenylyltransferase) gene as screening marker, two chloroplast transformation vectors of pTC2 harboring phaC2 gene only and pTGC harboring both phaC and phaG genes were constructed and introduced into to- bacco chloroplast genome through particle bombardment. PCR and Southern blot analysis confirmed the insertion of the introduced genes into chloroplast genome. The content of mcl-PHAs accumulated in transgenic plants was analyzed by gas chromatography, mcl-PHAs accumulated up to 4.8 mg/g dry weight (dw) in transgenic line S4-3; their monomers were 3-hydroxyoctanoate and 3-hydroxydecanoate. Accumulation of mcl-PHAs polymers in the tobacco chloroplast was also observed by transmission electron microscopy. To our knowledge, this is the first report on the synthesis of mcl- PHAs in tobacco via chloroplast genetic engineering.Medium-chain-length-polyhydroxyalkanoates (mcl-PHAs) belong to the group of microbial polyesters containing monomers ranging from 6 to 14 carbons in length. The key enzymes of their biosynthesis are PHA-polymerase (product of phaC gene) and 3-hydroxyacyl-acyl carrier protein-CoA transferase (product of phaG gene). With aadA (aminoglycoside 3'-adenylyltransferase) gene as screening marker, two chloroplast transformation vectors of pTC2 harboring phaC2 gene only and pTGC harboring both phaC and phaG genes were constructed and introduced into tobacco chloroplast genome through particle bombardment. PCR and Southern blot analysis confirmed the insertion of the introduced genes into chloroplast genome. The content of mcl-PHAs accumulated in transgenic plants was analyzed by gas chromatography, mcl-PHAs accumulated up to 4.8 mg/g dry weight (dw) in transgenic line S4-3; their monomers were 3-hydroxyoctanoate and 3-hydroxydecanoate. Accumulation of mcl-PHAs polymers in the tobacco chloroplast was also observed by transmission electron microscopy. To our knowledge, this is the first report on the synthesis of mcl-PHAs in tobacco via chloroplast genetic engineering.

关 键 词：媒介链长度 烟草 叶绿体 遗传工程 生物合成 

分 类 号：S572[农业科学—烟草工业]