应用改良3′ cDNA末端快速扩增PCR法（rapid amplification of cDNA ends，RACE）筛选扇贝抗菌肽基因。方法 设计M13和T7启动子分别修饰的基因特异性引物（gene special primer，GSP）、锚定引物，提取扇贝血淋巴总RNA进行改良3′ RACE和5′ RACE，获得cDNA准全长序列并在GenBank中进行同源性比较。结果 从扇贝血淋巴中获得3个cDNA的3′端序列基因和1个准全长cDNA（544 bp），在GenBank中未发现与之高度同源的基因。结论 从扇贝血淋巴中获得的1个准全长cDNA、3个cDNA的3′端序列可能属于新基因。应用改良3′ RACE能钓取含抗菌肽保守序列的新基因片段，该方法可行。

To screen antimicrobial peptide genes from scallop with improved PCR method of 3′ rapid amplification of cDNA ends (RACE). Methods Gene special primers and anchor primers modified by promoters M13 and T7 were designed. Total RNA was extracted from scallop and amplified by improved PCR method of 3′ RACE and 5′ RACE to obtain the full-length cDNA of antimicrobial peptide genes. The homology of the sub-full-length cDNA sequence was blasted in GenBank. Results Three 3′ end cDNA sequences and one sub-full-length cDNA (544 bp) were obtained. There was no similar gene in homology in GenBank. Conclusion One sub-full-length cDNA and three cDNAs 3′ end sequences from scallop hemolymph may be new genes. It is available to use improved PCR method of RACE to screen gene fragments containing antimicrobial peptide conserved sequence.

广东省自然科学基金资助项目（5011588）