Cloning and Identification of the cDNA and Genomic DNA Sequences of the Defensin Gene of Anopheles sinensis / 中国寄生虫学与寄生虫病杂志

ABSTRACT

Objective To clone and identify the cDNA sequence and genomic DNA sequence of Anopheles sinensis defensin gene. Methods Referring to the published defensin gene sequences of Aedes aegypti and Anopheles gambiae, pairs of primers were designed to amplify the cDNA sequence and genomic DNA sequence of Anopheles sinensis defensin gene with template of Anopheles sinensis total RNA by RT-PCR and genomic DNA by nested PCR, respectively. These amplified fractions were cloned and sequenced, and were further identified and analyzed by relevant bioinformatics softwares. Results Complete genomic DNA sequence was cloned, including 5′ and 3′ UTR fraction and two exons separated by a 85 bp intron. Besides, the whole cDNA sequence of Anopheles sinensis defensin gene with 324 bp was also cloned, and its ORF encoded 107 amino acids. The mature peptide had 40 amino acids residues. Conclusion The whole cDNA sequence and complete genomic DNA sequence of the defensin gene of Anopheles sinensis have been cloned and identified for the first time.