Src family kinases (SFKs), a class of non-receptor tyrosine kinases, mediate a wide aspect of cellular signaling pathways that regulate cell proliferation, differentiation, motility and survival. In this study, we identified and characterized for the first time a novel SFK homologue from Litopenaeus vannamei (designated as LvSrc). Sequence analysis showed that LvSrc had a high homology with the identified SFKs, especially those from invertebrates. LvSrc contained the conserved SH3, SH2 and tyrosine kinase domains, as well as the potential phosphorylation and lipid modification sites. Immunofluorescence analysis demonstrated that LvSrc was mostly localized at the plasma membrane and partly resided in the perinuclear vesicle and nucleus or whole cell. Infection with white spot syndrome virus (WSSV) could up-regulate the transcription and expression levels of LvSrc and further induced its phosphorylation, suggesting that LvSrc was implicated in WSSV infection. Furthermore, our co-immunoprecipitation result confirmed the interaction between Src and focal adhesion kinase (FAK) in shrimp, while the phosphorylation of FAK was markedly enhanced by co-expression with LvSrc. In sum, our studies suggested that LvSrc might act in the FAK-regulated signaling pathway during WSSV infection, which would give us a better insight in understanding the role of SKFs in host-virus interactions in crustaceans.