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《Molecular Plant Breeding》 2003-01
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Construction of Two Gene Plant Expression Vector Harboring phaC1、phaC2 Genes and Obtainment of Transgenic Tobacco

Zheng Junrong 1, 2 Zhou Peng 1 Hong Kui 1* 1 State Key Biotechnology Lab for Tropical Crops, China Academy of Tropical Agriculture Science, Haikou, 571101 2 Engineering School, South China University of Tropical Agriculture, Danzhou, 571737  
phaC1 and phaC2, two key polyhydroxyalkanoates polymerase genes of PHA biosynthesis were amplified and cloned from chromosomal DNA of Pseudomonas psuedoalcaligenese YS1 using PCR. At the same time, phaC1 gene was modified using PCR. Three plant expression vectors: pC3C1 (containing phaC1), pC3C1 (containing phaC2), pC3C1C2 (containing phaC1 and phaC2) were constructed. The high performance two gene expression vector pC3C1C2 was transformed to Nicotiana tobacum Honghuadajinyuan plants mediated by Agrobacterium tumefaciens EHA105. A number of Kanamycin resistant transformants were obtained two months later. They were morphologically normal, but grew slower comparing with the control tobacco. 32% transformants were stable integration of phaC1 and phaC2 confirmed by PCR and PCR Southern. Moreover, extracting and purifying PHAs directly from positive transformants by chloroform hypochloric acid showed that 25 6%of transformed tobacco plants could produce PHAs.
【CateGory Index】: Q943.2
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