應(yīng)用改良3′ cDNA末端快速擴(kuò)增PCR法（rapid amplification of cDNA ends，RACE）篩選扇貝抗菌肽基因。方法 設(shè)計(jì)M13和T7啟動(dòng)子分別修飾的基因特異性引物（gene special primer，GSP）、錨定引物，提取扇貝血淋巴總RNA進(jìn)行改良3′ RACE和5′ RACE，獲得cDNA準(zhǔn)全長序列并在GenBank中進(jìn)行同源性比較。結(jié)果 從扇貝血淋巴中獲得3個(gè)cDNA的3′端序列基因和1個(gè)準(zhǔn)全長cDNA（544 bp），在GenBank中未發(fā)現(xiàn)與之高度同源的基因。結(jié)論 從扇貝血淋巴中獲得的1個(gè)準(zhǔn)全長cDNA、3個(gè)cDNA的3′端序列可能屬于新基因。應(yīng)用改良3′ RACE能釣取含抗菌肽保守序列的新基因片段，該方法可行。

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.

廣東省自然科學(xué)基金資助項(xiàng)目（5011588）