Drosophila SMART testThe SMART was

Drosophila SMART test

The SMART was essentially performed as described by Graf et al. [20] For this assay, the following cross of Drosophila melanogasterflies was used: ORR (i); ORR (ii); flr3/In (3LR) TM3, BdS virgin females were crossed with mwh males (flies that were kindly provided by Agarkar Institute, Pune). The first strain is characterized by constitutively high cytochrome P-450 activity. The markers mwh and flr3 (misshapen, flr-like hairs) are recessive wing-hair mutations located on the third chromosome at 0.3 and 38.8, respectively. This test is able to detect a wide spectrum of genetic alterations including point mutations, deletions, unbalanced half-translocation and mitotic recombination, chromosomal loss, and non-disjunction as described in Graf et al. [20]

Transheterozygous larvae were obtained by parental crosses between flr3 virgin females and mwh males. Eggs were collected from this cross during 8-h period in culture bottles containing fresh standard Drosophila medium (wheat powder, jaggery, agar agar, propionic acid, and water cooked). After 72 h, third instar larva were floated off with tap water and transferred to plastic vials containing 1.5 g of Drosophila instant medium rehydrated with 9 mL of freshly prepared test solutions (mutagens, mutagens plus extracts, distilled water, and MMS used at positive control at 0.1 and 0.05 mM). For each treatment group in a total of 4,000 larvae, 200 in each vial were employed. The larvae were fed on this medium until pupation of the surviving larvae. All the experiments werecarried out at 24 ± 1°C and at ~60% relative humidity.

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Uji cerdas DrosophilaSMART pada dasarnya dilakukan seperti yang dijelaskan oleh Graf et al. [20] untuk assay ini, salib berikut Drosophila melanogasterflies digunakan: ORR (i); ORR (ii); flr3/TM3 (3LR), BdS perawan Perempuan menyilang dengan laki-laki mwh (lalat yang ramah disediakan oleh Agarkar Institute, Pune). Ketegangan pertama ini ditandai dengan constitutively tinggi sitokrom kegiatan P-450. Penanda mwh dan flr3 (Cacat, flr-seperti rambut) adalah mutasi-mutasi resesif sayap-rambut yang terletak pada kromosom ketiga 0.3 dan 38.8, masing-masing. Tes ini mampu mendeteksi spektrum yang luas dari perubahan genetik yang termasuk titik mutasi-mutasi, penghapusan, tidak seimbang rekombinasi setengah-translokasi dan mitosis, kehilangan kromosom, dan bebas-kesenjangan seperti yang dijelaskan di Graf et al. [20]Transheterozygous larvae were obtained by parental crosses between flr3 virgin females and mwh males. Eggs were collected from this cross during 8-h period in culture bottles containing fresh standard Drosophila medium (wheat powder, jaggery, agar agar, propionic acid, and water cooked). After 72 h, third instar larva were floated off with tap water and transferred to plastic vials containing 1.5 g of Drosophila instant medium rehydrated with 9 mL of freshly prepared test solutions (mutagens, mutagens plus extracts, distilled water, and MMS used at positive control at 0.1 and 0.05 mM). For each treatment group in a total of 4,000 larvae, 200 in each vial were employed. The larvae were fed on this medium until pupation of the surviving larvae. All the experiments werecarried out at 24 ± 1°C and at ~60% relative humidity.