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    • 7. 发明授权
    • Method and materials for bisulfite conversion of cytosine to uracil
    • 亚硫酸氢盐转化为尿嘧啶的方法和材料
    • US07371526B2
    • 2008-05-13
    • US10926531
    • 2004-08-26
    • Gerald ZonVictoria L. Boyd
    • Gerald ZonVictoria L. Boyd
    • C12Q1/68G01N33/53C07H21/00
    • C07H21/02C07H21/04C12Q1/6886C12Q2600/154Y10S435/81Y10S436/808
    • The invention provides methods and materials for conversion of cytosine to uracil. In some embodiments, a nucleic acid, such as gDNA, is reacted with at least one bisulfite salt having the formula X+HSO3− or Y+2(HSO331)2; wherein X+ is ammonium ion, a tetraalkyl ammonium ion, or a group 1A ion other than sodium; and Y+2 is a group 2A ion or a group 7B ion; under conditions effective to convert at least one cytosine nucleobase to a uracil nucleobase. In some embodiments, X+ comprises at least one of lithium ion, potassium ion, ammonium ion, tetraalkylammonium ion, magnesium ion, manganese ion and calcium ion. In some embodiments, the reacting is performed optionally in the presence of a polyamine catalyst and/or a quaternary amine catalyst. Also provided are kits that can be used to carry out methods of the invention.
    • 本发明提供了将胞嘧啶转化成尿嘧啶的方法和材料。 在一些实施方案中,核酸,例如gDNA,与至少一种具有下式的亚硫酸氢盐反应:具有式X + 3,...,..., 2 + 2(HSO 3 3)3)2。 其中X +为铵离子,四烷基铵离子或除钠以外的1A族离子; Y 2 + 2是2A族离子或7B族离子; 在有效将至少一个胞嘧啶核碱基转化为尿嘧啶核碱基的条件下。 在一些实施方案中,X + +包括锂离子,钾离子,铵离子,四烷基铵离子,镁离子,锰离子和钙离子中的至少一种。 在一些实施方案中,反应任选在多胺催化剂和/或季胺催化剂的存在下进行。 还提供了可用于实施本发明方法的试剂盒。
    • 8. 发明授权
    • Thiohydantoin formation and selective modification of the carboxy
terminus of an aspartic acid- and/or glutamic acid-containing protein
    • 硫代海因素形成和天冬氨酸和/或谷氨酸的蛋白质的羧基末端的选择性修饰
    • US5665603A
    • 1997-09-09
    • US272496
    • 1994-07-08
    • Victoria L. BoydMeriLisa BozziniRobert J. DeFranco
    • Victoria L. BoydMeriLisa BozziniRobert J. DeFranco
    • C07K1/12G01N33/68A61K38/02
    • G01N33/6842C07K1/12G01N33/6821
    • A method of forming a thiohydantoin from an N-protected amino acid is described. The method employs a phosphate compound selected from the group consisting of(R.sub.1 O)(R.sub.2 O)P(.dbd.O)X and(R.sub.1 O)(R.sub.2 O)P(.dbd.O)--O--P(.dbd.O)(OR.sub.3)(OR.sub.4)to form acylphosphate moieties from the carboxyl groups of internal aspartic acid and glutamic acid residues and an acylphosphate moiety at a C-terminal carboxyl. The later acylphosphate, unlike the internal acylphosphates, spontaneously cyclizes to an oxazolone, which is less reactive with nucleophilic reagents. R.sub.1 and R.sub.2 are each alkyl, aryl, or alkaryl groups which are the same or different and which may be covalently linked to each other; R.sub.3 and R.sub.4 are each alkyl, aryl, or alkaryl groups which are the same or different and which may be covalently linked to each other; and X is a leaving group, such as chlorine or bromine, which is substantially unreactive towards thiohydantoins. The acylphosphate and oxazolone moieties are then reacted with a thiocyanate reagent under conditions effective to convert the internal acylphosphates to amides and the terminal oxazolone to thiohydantoin, thereby permitting selective C-terminal thiodantionation of aspartic acid- and/or glutamic acid-containing proteins.
    • 描述了从N-保护的氨基酸形成硫代乙内酰脲的方法。 该方法使用选自(R1O)(R2O)P(= O)X和(R1O)(R2O)P(= O)-OP(= O)(OR3)(OR4))的磷酸酯化合物 内部天冬氨酸和谷氨酸残基的羧基的酰基磷酸酯部分和C末端羧基上的酰基磷酸酯部分。 后来的酰基磷酸酯与内酰基磷酸酯不同,自发环化成与亲核试剂反应性较低的恶唑酮。 R1和R2各自是相同或不同的并且可以彼此共价连接的烷基,芳基或烷芳基; R 3和R 4各自是相同或不同的并且可以彼此共价连接的烷基,芳基或烷芳基; 并且X是对硫代乙内酰脲基本上不反应的离去基团,例如氯或溴。 然后将酰基磷酸酯和恶唑酮部分与硫氰酸酯试剂在有效将内部酰基磷酸酯转化为酰胺,并将末端恶唑酮转化为硫代乙内酰脲的条件下反应,从而允许含有天冬氨酸和/或谷氨酸的蛋白质的选择性C-末端硫代去离子。