对虾传染性皮下及造血组织坏死病毒(福建株)基因组的克隆

Genome cloning of the infectious hypodermal and hematopoietic necrosis virus(IHHNV) isolated from Fujian,China

对虾传染性皮下及造血组织坏死病毒(IHHNV)是世界各地养殖和野生对虾的重要病原之一.目前,GenBank已收录了分离自夏威夷、加利福尼亚、泰国、中国台湾等地的IHHNV全长或部分基因组序列.虽然中国大陆在养殖对虾中也普遍检测到了IHHNV,但未见其基因组序列报道.本文首先利用DNA末端加尾和嵌套PCR克隆了IHHNV基因组两末端序列,并根据测定的末端序列设计引物,PCR扩增出中国福建株IHHNV基因组序列.

Infectious hypodermal and hematopoietic necrosis virus （IHHNV） is one of important pathogens causing farmed and wild shrimp diseases. At present, the complete or partial IHHNV genomic sequences, which were isolated from Hawaii, California, Thailand and Taiwan in China, have been deposited in GenBank. Although the IHHNV-infected penaeid shrimps are detected often in China's Mainland, no IHHNV genome sequence from Chinese isolate was reported. In this experiment, the IHHNV genome ends were cloned and sequenced by means adding poly （A） to the 3＇ end of the DNA and nested PCR. According to sequences of the 5＇ and 3＇ ends termini, the fulllength IHHNV genome was amplified by PCR with specific primers and sequenced.