Bovine viral diarrhea virus (BVDV) belongs to the Pestivirus genus of the Flaviviridae family and has worldwide distribution, being one of the main causes of economic losses in cattle raising. The genome of pestiviruses is a single strand of positive-sense RNA with a length of 12.3 kb, which encodes one open reading frame flanked by untranslated regions. E2 glycoprotein is required for binding to cell-surface receptors and it also contains major antigenic determinants. The nucleotide sequence coding E2 is the most variable part of the viral genome. The heterogeneity that exists among circulating strains causes problems in the development of effective vaccines and reliable diagnostics. In this study, and for the first time analysis was made of the E2 glycoprotein coding sequences of 14 Polish BVDV-1 strains which belong to four subtypes: 1b (n = 7), 1f (n = 3), 1s (n = 3), and 1r (n = 1). These sequences showed evidence of strong purifying (negative) selection. However, we also identified positively selected sites. The availability of E2 sequences of Polish BVDV strains for reference, knowledge gained through epitope prediction attempts, and information on protein glycosylation sites can afford a better understanding of host–pathogen interactions.