Aim: Anautogenous mosquitoes require a blood meal before they can lay their first batch of eggs. Unluckily, this happens at the consequence of pathogen transmission and the spread of vector-borne diseases. The reproductive success of all oviparous species including insects depends on vitellogenin (Vg) biosynthesis and its accumulation in the developing oocytes. Therefore, during the present study, the temporal and spatial expression of the Vg gene has been studied in Anopheles culicifacies, which is the major vector in Indian subcontinent. Methodology: The Dhera strain of An. culicifacies Species A was maintained in an insectary at appropriate rearing conditions. The RNA was extracted from the female fat bodies after blood feeding at different time intervals. The temporal and spatial expression patterns of the Vg gene was examined by Real-time PCR using SYBR Green dye. Result: The An. culicifacies vitellogenin (AncuVg) showed significant and strong expression in fat body tissues only as compared to other tissues at specific time interval after blood feeding. Time and tissue specific expression of the gene showed induction of the gene at 3 h PBM and peak expression was observed after 24 h of blood feeding. Discussion: The peak expression of Vg gene observed at 24h corresponds to the peak levels observed earlier for 20-E after 24h of blood meal and also corresponds with the timing of ookinete invasion and oocyst formation. Conclusion: The molecular events of vitellogenesis as observed in case of An. culicifacies, may prove beneficial to develop novel control strategies to combat parasite transmission because the timing of Vg expression coincides with ookinetes invasion in general.