Biochem Lit Essay Example | Topics and Well Written Essays - 500 words

Biochem Lit - Essay Example It was ascertained in this course of study that ChiB, a gene coding for endo Chitinase B, is in part responsible for induction of autolysis by flbA gene. Also though ChiB was found to be necessary for ∆flbA induced autolysis its overexpression did not accelerate or enhance hyphal disintegration thus suggesting that ChiB is necessary but not sufficient for autolysis in submerge-cultured A.nidulans. Alamar Blue reduction test which corresponds to the activity of the living cell mitochondria was employed to assess the dispensability of ChiB gene in autolysis. By AB reduction it was seen that both Wild Type strain and ChiB deletion ∆ChiB mutant showed the same death pattern from day 1 to day 8. Thus though ChiB was required for autolysis it was not totally essential. To study the proteins involved in autolysis, the proteins upregulated in ∆flbA and ∆flbA∆ChiB were studied and five proteins were found to be upregulated. These were identified as chitinase (Chi B), dipeptidyl peptidase V (DppV), O-glycosyl compound hydrolase, -N-acetylhexosaminidase (NagA), and myo-inositol-1-phosphate synthase (InoB) on SDS PAGE after Liquid chromatography-tandem mass spectroscopy. Off these 5 proteing NagA was speculated to be associated with cell death. This was because ∆NagA mutant showed a significantly higher percentage reduction in Alamar Blue as compared to other proteins on day 2 of incubation indicating more viable cells. The absence of InoB or AN2395.2 (a probable hydrolase) was seen to cause accelerated cell death and disorganization of mycelial pellets suggesting that these proteins might be needed for the controlled progression of cell death. FlbA gene negatively controls G protein mediated proliferation signaling likely by acting as GTPase activating protein for FadAGÃŽ ±. It was also concluded in other studies that FadA and SfaD signaling negatively regulate the cell wall chitin content and cell wall structure in A.nidulans. FadA was also found to mediate