The purpose of this study is to explore how the relation between gender and humour is maintained and challenged in the Swedish TV show Parlamentet. Using an interactional approach based on Judith Baxter’s theoretical framework for feminist post-structuralist discourse analysis, the study focuses on how communicative strategies are used in negotiating for the ideal position of “the successful comedian”. This position is seen as an ideal position in Parlamentet, where comedians do political parody. In the conversation sequences examined in this study, both verbal and bodily semiotic modes have been analyzed using conversation analysis and multimodal analysis. The study focuses on how different communicative strategies encourage or reject the comedians and how this leads to different possibilities in negotiating for a powerful position, the position of the successful comedian. Humour is culturally seen as a male-coded discourse. The relations between humour and gender that are in focus are the hegemonic notions of women as lacking a sense of humour or being less humorous than men. The notions of gender are based on Judith Butler’s theories and hence seen as something performed through discourse and within a rigid regulatory frame where the subject’s possibilities are not infinite. The results of the study show that the female and male comedians do not have the same possibilities in the negotiation for the position of “the successful comedian”. The male comedians are strongly encouraged to a greater extent than the female comedians. The female comedians are also rejected to a larger extent than the male comedians. The male comedians are mostly rejected by the moderator whereas the female comedians are rejected by the moderator, male comedians and female comedians. The female comedians encourage other comedians to a larger degree than the male comedians. A significant finding of this study is that the relations between humour and gender, where women are seen as less humorous than men, are maintained when female comedians through discourse are made less humorous. However, there are strategies in the interaction that do encourage the female comedians or that lead to female comedian’s resist being rejected. These strategies indicate that the hegemonic cultural notions of women as less humorous than men are also challenged in this public discourse.