Assays of PCB congeners and organochlorine insecticides with the transgenicArabidopsisand tobacco plants carrying recombinant guinea pig AhR and GUS reporter genes

The transformation of coffee plantlets with the cry1ac gene of Bacillus thuringiensis was achieved by biolistic using either the whole pUBC plasmid or only the ubi-cry1ac-nos genetic cassette. The cry1ac gene was inserted into coffee plants in order to confer resistance to the leaf miner Leucoptera coffeella, an insect responsible for considerable losses in coffee crops. Bearing in mind that the genetic cassettes used for this study lack reporter genes and/or selection marker genes, the parameters for the transformation procedure by biolistic were previously standardised with a plasmid carrying the gus reporter gene. The presence of the cry1ac gene in young plantlet tissues was determined by PCR, Southern blot and reverse transcription-PCR. Our results show that the obtainment of viable coffee plantlets, transformed by bombardment with the cry1ac gene and without selection markers nor reporter genes, is feasible.

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Effect of nuclear matrix attachment regions on transgene expression in tobacco plants.

Acta Biochimica Polonica ◽

10.18388/abp.2001_3898 ◽

2001 ◽

Vol 48 (3) ◽

pp. 637-646 ◽

Cited By ~ 16

Author(s):

W Nowak ◽

M Gawłowska ◽

A Jarmołowski ◽

J Augustyniak

Keyword(s):

Transgenic Plants ◽

Reporter Gene ◽

Transgene Expression ◽

Binary Vector ◽

Special Property ◽

Matrix Attachment Regions ◽

Plant Populations ◽

Tobacco Plants ◽

Gus Reporter ◽

Loop Domains

Matrix attachment regions (MARs) are thought to participate in the organization and segregation of independent chromosomal loop domains. Although there are several reports on the action of natural MARs in the context of heterologous genes in transgenic plants, in our study we tested a synthetic MAR (sMAR) with the special property of unpairing when under superhelical strain, for its effect on reporter gene expression in tobacco plants. The synthetic MAR was a multimer of a short sequence from the MAR 3' end of the immunoglobulin heavy chain (IgH) enhancer. This sMAR sequence was used to flank the beta-glucuronidase (GUS) reporter gene within the T-DNA of the binary vector pBI121. Vectors with or without the sMARs were then used to transform tobacco plants by Agrobacterium tumefaciens. Transgenic plants containing the sMAR sequences flanking the GUS gene exhibited higher levels of transgene expression compared with transgenic plants which lacked the sMARs. This effect was observed independently of the position of the sMAR at the 5' side of the reporter gene. However, variation of the detected transgene expression was significant in all transformed plant populations, irrespective of the construct used.

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Multiresidue Method for Determination of Organochlorine Insecticides and Polychlorinated Biphenyl Congeners in Fatty Processed Foods

Journal of AOAC International ◽

10.1093/jaoac/79.6.1434 ◽

1996 ◽

Vol 79 (6) ◽

pp. 1434-1446 ◽

Cited By ~ 17

Author(s):

Anna Sannino ◽

Paola Mambriani ◽

Mirella Bandini ◽

Luciana Bolzoni

Keyword(s):

Polychlorinated Biphenyl ◽

Methylene Chloride ◽

Gas Chromatography Mass Spectrometry ◽

Animal Origin ◽

Electron Capture Detection ◽

Matrix Interference ◽

Selected Ion Monitoring ◽

Pcb Congeners ◽

Organochlorine Insecticides

Abstract A previously published gel permeation chromatographic (GPC) procedure for determination of organophosphorous insecticides is used to determine 24 organochlorine (OC) pesticide and 9 polychlorinated biphenyl (PCB) residues in 8 fatty preserved foods of vegetable and animal origin. Samples are extracted with methylene chloride and cleaned up by automated GPC with a Biobeads SX3 column and a methylene chloride–cyclohexane (15 + 85) eluant. Compounds are quantitated by gas chromatography/mass spectrometry (GC/MS) with selected ion monitoring using a DB-5 column. Average recoveries of OC pesticides from samples fortified at 0.010–0.080 mg/kg ranged from 73 to 98%. Average recoveries of 9 PCB congeners spiked at 2 levels (0.0025 and 0.020 mg/kg) were between 85 and 104%. Quantitation limits were between 0.002 and 0.025 mg/kg, depending on the compound. The purified extracts were analyzed further by GC with electron capture detection (ECD), and the results were compared with those obtained by MS. Determination of some OC pesticides and PCB congeners was not possible by ECD because of matrix interference.

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