Expression Pattern of Lipase Gene Family LIP 1 10 of Candida Albicans under Different Physiological Conditions Inducing Hyphae

Abstract

Candida albicans proved to be the most frequent opportunistic pathogen associated with life threatening infections, whose proliferation is regulated by the Lipase (LIP) gene family. C.albicans exhibit ten members in Lipase (LIP1-10) gene family that encodes ten hydrolytic enzymes involved in invasion to host cells. The microscopic observation of control and test samples after 6 hrs of incubation shows a greater hyphae development in the flasks added with inducers as compare to control samples. Tween 80 was used as a lipase substrate for all induction systems, which showed a demarcated zone of intensification with increasing incubation time. Evaluation of differential gene expression patterns of the LIP (LIP1-10) gene family under the hyphae form induced by two environmental and nutritional conditions during different time intervals using Real-Time qPCR. Different inducers like Temperature, pH, GlcNAc and Proline were used to find the temporal expression as compared to control conditions of 30°C and pH 6.5. GAPDH housekeeping gene was used as reference in expression conditions. The integrity and concentration of extracted RNA was confirmed by Agarose Gel electrophoresis. Differential expressions of the LIP genes were found at control and test temperature, pH, GlcNAc and Proline. LIP1-3 were highly active after giving hyphae-inducing elicitors for the first 90 minutes of exposure. LIP1, LIP2, LIP3, LIP5, LIP8 and LIP10 were highly modulated in test conditions in different incubation time. The rest of the LIP alleles expressions at incubation times after inducing the hyphae by elicitors. Clinically, it is difficult to control LIP gene family expression at different pH and temperatures with GlcNAc and Proline. In silico study involving physicochemical, phylogenetic, structural, molecular modeling and docking Studies of LIP genes and proteins family in correlation with RT-qPCR results. A variation in the LIP genes and proteins sequences under different micro-environments can reveal structural and evolutionary changes