Gobiocypris rarus is a unique species of small fish in China. At present study, a fragment of 6 299 bp was cloned by PCR method. It was found that the sequence identity of coding area of this fragment is 100% compared to the cDNA sequence of vitellogenin of Gobiocypris rarus. By using genome walking technique, a product of 1 285 bp was acquired based on the 6 299 bp sequence. Finally, a sequence of 7 573 bp was gotten by comparing the two fragments of 1 285 bp and 6 299 bp, with a vitellogenin gene of 6 595 bp and upstream sequence of 978 bp. After the ORF analysis , it was found there were 26 exons and 25 introns, the CDS was 3 900 bp which code 1 200 amino acid. Blast analysis found there existed 80% sequence identity between the upstream fragment of 220-838 bp of our sequence and the promoter sequence of 2 875-3 469 bp of vitellogenin of Acheilognathas yamatsutae.