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マツカワのウイルス性神経壊死症原因ウイルス遺伝子の検出に及ぼすPCR条件の検討

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Title: マツカワのウイルス性神経壊死症原因ウイルス遺伝子の検出に及ぼすPCR条件の検討
Other Titles: Revelation of effective methods for detection of viral nervous necrosis virus gene using polymerase chain reaction in barfin flounder, Verasper moseri
Authors: 渡辺, 研一1 Browse this author
吉水, 守2 Browse this author →KAKEN DB
Authors(alt): Watanabe, Ken-ichi1
Yoshimizu, Mamoru2
Keywords: Barfin flounder
PCR
Viral nervous necrosis
Viral detection
Issue Date: 20-Mar-2001
Publisher: 日本水産増殖学会
Journal Title: 水産増殖
Volume: 49
Issue: 1
Start Page: 85
End Page: 90
Publisher DOI: 10.11233/aquaculturesci1953.49.85
Abstract: Detection rate of viral nervous necrosis (VNN) virus gene using polymerase chain reaction was investigated. Tested specimens were: just hatched larvae, heads of larvae, eye or brain of juveniles, ovarian fluid and sperm obtained from brood fish. The specimens were mixed with a 10-fold serial dilution of virus solutions prepared from the eyes and brain of the Pacific Cod, Gadus macrocephalus, affected with VNN. For nucleic acid extraction, a comparison was made between 20-proteinase K, SDS- proteinase K, acid guanidium phenol chloroform, Isogen, TRIzol, RNA isolation kit, Catrimox-14, and High Pure Viral Nucleic Acid Kit. Isogen and/or RNA isolation kit showed the highest detection rate. PTC-200 and PJ 480 thermal cyclers were more effective than the PC-700 model. In comparison of reverse transcriptase, AMV, M-MLV, and Super Script II were tested; r Taq or Ex Taq was used as the DNA polymerase. Pairing of Super Script and Ex Taq was most effective. In PCR programs, 3-temperature PCR was more effective than 2-temperature PCR.
マツカワを材料とした場合の,核酸抽出法,PCR反応温度,反応酵素,機器等のウイルス性神経壊死症原因ウイルス遺伝子の検出に及ぼす影響について比較検討した。核酸抽出法としてはIsogenおよびRNA Isolation Kitが,cDNAの増幅に際して変性,アニーリング,伸長反応を3種類の反応温度により行う方法が,逆転写酵素としてはSuper Script II,DNAポリメラーゼはEx Taqが,サーマルサイクラーはPTC-200およびPJ480が優れていた。
Rights: © 2001 日本水産増殖学会
Type: article
URI: http://hdl.handle.net/2115/53095
Appears in Collections:水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 吉水 守

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