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In order to independently validate the transcriptomic responses of the genes found to be responsive to heat and cold stress, 16 rice genes and their respective Arabidopsis orthologues (32 genes in total) were analysed by qRT-PCR.
Among the responsive proteins, the majority of them were associated exclusively with water-deficit stress, and those proteins responsive to heat stress were responsive to water-deficit stress also.
Because the transcriptional profile of larvae greatly differed from cells, we investigated whether genes responsive to heat shock in larvae were any more likely to be associated with HSF.
A qPCR analysis showed that MaMSD1A was responsive to heat, drought and salt stress, while MaMSD1B was responsive to heat and drought stress.
MaMSD1C was only responsive to heat stress, and MaMSD1D was not responsive to any stress.
In our study, 106 of 178 MYB genes were detected as responsive to heat stress.
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Among these are several cDNAs encoding putative chaperones, which we have demonstrated to be responsive to heat-shock.
A YFP/SV40pA cassette was cloned downstream of a 4xUAS/dHSP70 element (non responsive to heat-shock; kind gift of M. Gonzalez-Gaitan).
Many genes responsive to heat- or cold-shock have been identified in S. aureus based on transcriptome data [ 91], but a systematic and comprehensive analysis of the fitness of temperature-sensitive or resistant mutants has not been reported.
SSH-identified transcripts (e.g. HSP90α, GRP94, GRP78, HSP70-1, HSP47) that were not only highly responsive to heat-shock, but also dysregulated in all three tissues studied, encode proteins that are known to prevent both programmed cell-death and aggregation.
Although HSP70s have been shown to be anti-apoptotic in sea bream (Sparus auratus) primary macrophage cultures [ 51], previous studies have reported that HSP70-1 proteis is not responsive to heat-stress in cod [ 29, 32].
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