The cyanobacterium Oscillatoria agardhii was isolated from the fresh water Mawatha lake, Jaipur and was grown in Zarrouk's medium at 25 ± 2°C, illuminated with white fluorescent light at the intensity of 2 500 lux with 12:12 h light and dark photoperiod. The effects of photoperiod and temperature on the growth and protein expression by Oscillatoria agardhii were studied under different controlled culture conditions (ALR, ALC, CLR, CLC, and NDL), measuring optical density, cell count and dry weight. Protein content was measured quantitatively by Bradford assay and qualitatively by SDS-PAGE. The densitometric analysis was also carried out for the measurement of the expression level of different proteins/peptides under different culture conditions. Maximum growth and protein content were observed in ALR condition while minimum was in the CLC. Alternate light and dark periods proved efficient as contrasting banding patterns were observed with many new unique polypeptides such as 32, 36.3, 47.9, 60.8, and 67.0 kDa, whereas, expressions of three polypeptides of 57.2, 110.1, and 117.3 kDa were inhibited in constant light cultures.