Gobiocypris rarus is a small cyprinids (carp) ，as a new model organism used in experimental research.In this study, using the ordinary PCR method to get a long sequence of 6299 bp. According to 5 'end of 6299 bp sequence , design two pairs of specific primers, using walking genome technology,to get 1285 bp sequence.7573 bp sequence after sequence assembly , which vitellogenin gene sequences of 6595 bp, upstream regulatory sequences of 978 bp. By open reading frame analysis , Gobiocypris rarus vitellogenin gene contains 26 exons and 25 introns, encoding 1299 amino acid sequence, CDS length 3900 bp. By BLAST comparison, Gobiocypris rarus vitellogenin upstream regulatory sequence 220 bp-838 bp interval and Acheilognathus yamatsutae vitellogenin gene promoter sequences 2857 bp-3144 bp interval have 80% similarity.