| 查看: 1876 | 回复: 4 | |||
[交流]
求助酶活测定方法 PAL、CHS、F3H、DFR、LDOX
|
|
谁能提供以下植物中 酶活性测定方法： PAL、CHS、F3H、DFR、LDOX 谢谢了！ |
回复此楼» 猜你喜欢
欢迎发来filecode的Mz6后的代码验证其规律
已经有70人回复
-
有时候，自然基金真的不能太认真 （我的申报经验）
已经有12人回复
-
招聘 金属平磨液，抛光液研发工程师
已经有3人回复
-
时间戳又变了8-15
已经有23人回复
-
filecode=后面第一个是大写字母
已经有7人回复
-
filecode
已经有12人回复
-
小木虫上这么多卖论文的，真有人买论文么？感觉没必要啊
已经有13人回复
-
哪位老哥知道今年的国自然具体哪一天放榜？
已经有7人回复
-
关于Filecode分析方法
已经有13人回复
-
各位道友，我要去昆明玩几天，回来见。
已经有8人回复
» 抢金币啦！回帖就可以得到:
-
祝福！基金中中中！！！！
+1/398
-
西安交大马伟/王亚洲课题组诚聘联培博士和硕士学生多名-有机生物电子学方向
+1/185
-
祝福！祈福！！基金中中中！
+5/85
-
中国科学院空天院精密干涉测量团队 精密机械结构工程师招聘启事
+1/83
-
中国科大-合肥国家实验室冷原子量子网络团队招聘启事
+2/56
-
广西大学 化学化工学院 新入职教师招生 化学、化学工程与技术、材料化工专业
+1/48
-
上海一恒贝茵公司全国均可联系销售或售后服务
+1/39
-
清华大学深圳国际研究生院招聘-博士后（长期有效）
+1/29
-
物理学报录用，散币感谢合作者和审稿人
+2/26
-
祈福
+1/21
-
希望这篇论文能中
+2/12
-
北京理工大学-集成电路与电子学院杰青团队-招博士后
+1/10
-
北理工集成电路杰青团队 | 诚招科助理
+1/8
-
关于Ghaffar-Parkins催化剂催化氰基水解为酰胺的问题
+1/6
-
南京医科大学-冯璋课题组-招收2027级生化与分子生物学专业博士和硕士研究生
+1/4
-
UTS 招收2027年入学博士生
+1/3
-
中国科学技术大学环境学院招聘招生
+1/2
-
海南大学化学院-科研助理招聘 （可读博，可与华中科技大学，天津大学联培）+博后招聘
+1/2
-
悉尼科技大学（QS 第87）数据科学/AI招收2027年秋季入学全奖及CSC博士生
+1/1
-
祈福今年中标教育部和省社科都中标，为退休倒计时画上圆满的句号
+1/1
2楼2014-09-15 14:32:02
★
小木虫: 金币+0.5, 给个红包，谢谢回帖
小木虫: 金币+0.5, 给个红包，谢谢回帖
|
这个好像很难，我查到了2种方法 第一种：ASSAY OF CHALCONE SYNTHASE ACTIVITY BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY，PhytochemistVryo,l . 34,N o. 5, pp. 1225-1229，1993 第二种： CHS[12] chaleone synthase，查儿酮合成酶 Chemicals, [2-14C]Malonyl-CoA (1.71GBq•mmo1-1) was purchased from N.E.N. (Boston, Mass., USA) and [U-14C]phenyl-alanine (16.65GBq•mmol-1) from Amersham (Braunschweig, Germany). 4-Coumaroyl-CoA was synthesized by Dr. P.A. Bäumker (Institut für Botanik, Münster, Germany; Bäumker et al. 1988)[13]. Enzyme preparations. ö Pbenylalanine ammonia-lyase: Leaves were cut into small pieces, frozen at -20°C and homogenized in a prechilled mortar in cold acetone (- 20°C The homogenate was filtered and the residue washed several times with cold acetone. The resulting dry powder was suspended in 0.1 M sodium borate buffer (pH 8.8) containing 5 mM dithioerythritol (DTE). Chalcone synthase. All steps were carried out at 0-4°C Leaf material was homogenized in 0.1 M potassium phosphate buffer (pH 6.8) containing 40 mM ascorbate, 1 mM EDTA, 10 mM thiourea硫脲, 5 mM DTE and 0.25 g Polyclar AT.(g FW)-1 (Serva, Heidelberg, Germany). The homogenate was centrifuged for 10 min at 10 000 g and the supernatant concentrated by ultrafiltration (Diaflo PM 30; Amicon, Witten, Germany). Enzyme assays, The activity of PAL was measured by the method of Amrhein and Zenk (1971). The activity of CHS was determined according to Beerhues and Wiermann (1988)[14], except that the pH of the incubation buffer was 7.8 and the final DTE concentration 0.5 mM. The analysis of the reaction products was performed as described by Lembach et al. (1989)[15]. (CHS activity was determined according to Beerhues and Wiermann (1985)[16]. The analysis of the reaction product was carried out using thin layer chromatography with silica gel plates and hexane: ethyl-acetate: methanol = 50:50:l (v/v/v) as a solvent system. For re-chromatography of the eluted product cellulose plates and 15% (v/v) ethanol were employed.) Protein determination. Protein concentration was measured according to the method of Bradford (1976), as modified by Read and Northcote (1981), using ovalbumin as a standard. Schöpker, H., M. Kneisel, L. Beerhues, et al. Phenylalanine ammonia-lyase and chalcone synthase in glands of Primula kewensis (W. Wats): immunofluorescence and immunogold localization. Planta, 1995. 196(4): 712-719. 13. PA, B., A. S and W. R. Metabolism of ferulic acid sucrose esters in anthers of Tulipa cv. Apeldoorn. II. Highly specific degradation of the esters by different esterase activities. Z. Naturforsch, 1988. 43c: 647-655. |
3楼2014-12-26 14:49:21
4楼2017-10-18 21:48:59
5楼2018-08-05 13:29:58
