Clone of C-type lectin gene of sea cucumber Apostichopus japonicus and optimization of experimental conditions using 3' RACE technique

The express sequence tag(EST) of C-type lectin of Apostichopus japonicus from GenBank was obtained.Based on this sequence,we utilize software of primer 5.0 to design a pair of RACE primer.After the PCR reaction,the cDNA sequence of C-type lectin of Apostichopus japonicus was obtained.Based on part of the known partial cDNA sequence,gene specific primers(GSPF) was designed which work together with universal primers(UPM).The 3' cDNA end of C-type lectin in Apostichopus japonicus was successfully cloned.At the same time,we optimized this experiment and an amplified fragment of 670 bp in length which overlapped the known C-type lectin sequence by 417 bp was obtained subsequently.The result showed that this sequence was consistent with expected aim gene.