Construction of Fosmid Genomic Library of Haliotis diversicolor Reeve and Screening of Hemocyanin Gene

The foot muscle of small abalone, Haliotis diversicolor was used to construct the Fosmid genomic library according to the CopyControl^TM Fosmid Library Production Kit. Fifty clones were randomly picked from library and identified by Not Ⅰ digestion. The positive rate was 100%. Lengths of these inserts were between 32 to 48 kb and with the average length 37.6 kb. Another 8 clones were randomly picked to do the end sequencing. With total 16 sequence results, 8 of them were sure from abalone genomes. Further more, specific primers were synthesized according to the 3′end sequence of hemocyanin gene of small abalone. With the PCR (Polymerase Chain Reaction) based screen method, a positive clone was obtained and identified as the hemocyanin type Ⅱ gene clone after Primer-Walking sequencings, Blast and phylogenetic tree analysis.