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    • 14. 发明授权
    • Method of multiplex microorganism detection
    • 多重微生物检测方法
    • US08298758B2
    • 2012-10-30
    • US10584393
    • 2004-12-24
    • Naoko HorikoshiSusumu KawasakiYukio OkadaKazuko TakeshitaTakashi SameshimaShinichi KawamotoKenji Isshiki
    • Naoko HorikoshiSusumu KawasakiYukio OkadaKazuko TakeshitaTakashi SameshimaShinichi KawamotoKenji Isshiki
    • C12Q1/68
    • C12Q1/686C12Q1/689Y02A50/451
    • The present invention is to provide a multiple detection method that can detect contaminating microorganisms existing in foods, including pathogenic Escherichia coli O157, Listeria monocytogenes and Salmonella spp., with high sensitivity comparable or even superior to official methods, comprising the steps of amplifying a plural number of target genes with a single PCR reaction tube and analyzing the same. The following steps are performed consecutively: (A) a step of extracting DNA of the target microorganisms to be detected by treating with at least a lytic enzyme such as Achromopepidase and Lysozyme and/or bacteriocin having lytic activity such as Enterolysine, a surfactant and a protein denaturing agent; and (B) a step of mixing a specific primer to the target microorganisms to be detected to perform multiplex PCR. Further, it is preferable to add a step of culturing with a culture condition where 1 CFU/100 g microorganisms becomes 10.sup.3 CFU/ml or more after 18 to 48 h of culture, for example that the pH after culture becomes 5.1 or more, before the step of extracting DNA of the target microorganisms to be detected.
    • 本发明提供一种可以检测存在于食品中的污染微生物的多重检测方法,包括致病性大肠杆菌O157,单核细胞增生李斯特氏菌和沙门氏菌,具有与官方方法相当或甚至优于其他方法的高灵敏度,包括以下步骤: 使用单个PCR反应管的目标基因数目并进行分析。 连续进行以下步骤:(A)通过用至少一种裂解酶如溶酶酶和溶菌酶和/或具有溶解活性的细菌素(例如肠溶素,表面活性剂和表面活性剂)进行处理来提取要检测的目标微生物的DNA的步骤 蛋白变性剂; 和(B)将特异性引物与要检测的目标微生物混合以进行多重PCR的步骤。 此外,优选在培养18〜48小时后,添加培养条件,其中1CFU / 100g微生物变成10μFCFU / ml以上,例如培养后的pH变为5.1 或更多,在提取要检测的目标微生物的DNA的步骤之前。
    • 15. 发明申请
    • METHOD FOR SELECTION OF HOP STRAIN, BLEEDING MARKER FOR USE IN SELECTION OF HOP STRAIN, AND PRIMER SET
    • 选择HOP菌株的方法,用于选择HOP菌株和PRIMER SET的BLEEDING MARKER
    • US20100055692A1
    • 2010-03-04
    • US12442116
    • 2007-09-19
    • Yukio OkadaKoichiro Koie
    • Yukio OkadaKoichiro Koie
    • C12Q1/68C07H21/04
    • C12Q1/6895A01H1/04C12Q2600/156
    • It is an object of the invention to screen for hop varieties with high α acid contents, as well as hop varieties with high contents of α acids, β acids, myrcene and/or xanthohumol in addition to α acids, within a short time period utilizing a molecular screening method that employs a breeding marker. The invention provides a breeding marker represented by the following (a) or (b), which is used for screening of hop varieties with high α acid contents.(a) A polynucleotide consisting of a nucleotide sequence of 20-1587 continuous nucleotides including nucleotide No. 899 of the polynucleotide consisting of the nucleotide sequence as set forth in SEQ ID NO: 5 (wherein the 899th nucleotide is t).(b) A polynucleotide consisting of the sequence complementary to the polynucleotide of (a) above.
    • 本发明的目的是筛选具有高α酸含量的啤酒花品种,以及具有高含量的α酸和啤酒花的啤酒花品种。 酸,月桂烯和/或黄曲霉酚以外的α酸,在短时间内利用采用育种标记的分子筛选方法。 本发明提供了以下(a)或(b)所示的育种标记,用于筛选具有高α酸含量的啤酒花品种。 (a)由包含SEQ ID NO:5所示核苷酸序列(其中第899位核苷酸为t)的多核苷酸的核苷酸序列号为899的连续核苷酸20-1587个核苷酸序列组成的多核苷酸。 (b)由与上述(a)的多核苷酸互补的序列组成的多核苷酸。
    • 18. 发明授权
    • Farnesyl pyrophosphate synthase protein, nucleic acid and promoter region thereof
    • 法呢基焦磷酸合酶蛋白,其核酸和启动子区
    • US06933374B2
    • 2005-08-23
    • US10148188
    • 2001-10-05
    • Yukio OkadaKazutoshi Ito
    • Yukio OkadaKazutoshi Ito
    • C12N9/10C12N15/60C12N15/82C07H21/04C12N15/00C12Q1/48
    • C12N9/1085C12N15/8225C12N15/8233C12N15/8243
    • The promoter region of the farnesyl pyrophosphate synthase gene that was expressed in the hop luplin gland in a specific manner was elucidated based on the genomic DNA of the hop farnesyl pyrophosphate synthase gene having the nucleotide sequence set forth in SEQ ID NO:2, the cDNA of the hop farnesyl pyrophosphate synthase gene having the nucleotide sequence set forth in SEQ ID NO:3, and the nucleotide sequence information on the genomic DNA and the cDNA. It will reveal the gene involved in the biosynthesis of secondary metabolites in a hop as well as the nucleotide sequence of the promoter gene that functions in the hop luplin gland in a tissue-specific manner. This will allow for the transformation of the hop by gene manipulations and the in vitro synthesis of the hop secondary metabolites.
    • 基于具有SEQ ID NO:2所示核苷酸序列的啤酒花法呢基焦磷酸合酶基因的基因组DNA,阐明了以特定方式在啤酒花叶绿体中表达的法呢基焦磷酸合酶基因的启动子区,cDNA 的具有SEQ ID NO:3所示核苷酸序列的啤酒花法呢基焦磷酸合酶基因,以及关于基因组DNA和cDNA的核苷酸序列信息。 它将揭示参与跳跃次生代谢产物的生物合成的基因以及以组织特异性方式在跳跃羽扇豆腺中起作用的启动子基因的核苷酸序列。 这将允许通过基因操作和跳跃次生代谢物的体外合成来转化啤酒花。