Tobacco Phospholipase D β1: Molecular Cloning and Biochemical Characterization

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Transgenic tobacco plants were developed containing a partial PLD clone in antisense orientation. The PLD isoform targeted by the insertion was identified. A PLD clone was isolated from a cDNA library using the partial PLD as a probe: Nt10B1 shares 92% identity with PLDβ1 from tomato but lacks the C2 domain. PCR analysis confirmed insertion of the antisense fragment into the plants: three introns distinguished the endogenous gene from the transgene. PLD activity was assayed in leaf homogenates in PLDβ/g conditions. When phosphatidylcholine was utilized as a substrate, no significant difference in transphosphatidylation activity was observed. However, there was a ... continued below

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Hodson, Jane E. December 2002.

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  • Hodson, Jane E.

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Transgenic tobacco plants were developed containing a partial PLD clone in antisense orientation. The PLD isoform targeted by the insertion was identified. A PLD clone was isolated from a cDNA library using the partial PLD as a probe: Nt10B1 shares 92% identity with PLDβ1 from tomato but lacks the C2 domain. PCR analysis confirmed insertion of the antisense fragment into the plants: three introns distinguished the endogenous gene from the transgene. PLD activity was assayed in leaf homogenates in PLDβ/g conditions. When phosphatidylcholine was utilized as a substrate, no significant difference in transphosphatidylation activity was observed. However, there was a reduction in NAPE hydrolysis in extracts of two transgenic plants. In one of these, a reduction in elicitor- induced PAL expression was also observed.

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  • December 2002

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  • Sept. 26, 2007, 2:56 a.m.

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  • Dec. 11, 2008, 5:01 p.m.

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Hodson, Jane E. Tobacco Phospholipase D β1: Molecular Cloning and Biochemical Characterization, thesis, December 2002; Denton, Texas. (digital.library.unt.edu/ark:/67531/metadc3341/: accessed November 19, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; .