LIBEST_028870
External Id
SAMN10259399
External Id
Nanyang Normal University
INSDC center name
2018-10-18T23:05:04.320Z
INSDC first public
2018-10-19T23:05:04.220Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
Partial cDNAs representing differentially expressed genes in Bt cells during the exposure to insect hemolymph were isolated by suppression subtractive hybridization, using a PCR select subtractive hybridization kit (Clontech, Palo Alto, CA, USA). First, exponentially growing Bt cells were exposed to a dilution of insect hemolymph and samples collected at 0, 2, 10 and 60 min postexposure to provide the cDNA pool for the 'tester' sample, whereas cDNA from unexposed 'same-age' Bt culture and extracted at the same time intervals was used as the 'driver' for the subtraction (cDNA library enriched in Bt genes that are up-regulated upon exposure to insect hemolymph). cDNA fragments were enriched with a 'suppression PCR' step prior to cloning (using pGEM-T Easy vector and JM109 E. coli competent cells). DNA sequencing of subtracted library clones with M13 forward and reverse primers was performed using an ABI377 automated sequencer (Applied Biosystems, Warrington, UK) and a BigDye terminator ready reaction kit (Perkin-Elmer Applied Biosystems, Foster City, CA, USA).
description
Bacillus thuringiensis serovar kurstaki str. HD-1
http://purl.obolibrary.org/obo/NCBITaxon_1261129
organism
kurstaki
serovar
HD-1
strain
Insect hemolymph-induced Bt genes
title