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Opti-MEMTM I Reduced Serum Medium

Opti-MEM TM I减少血清培养基

Company: Thermo Fisher Scientific
Catalog#: 31985-062
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Detection of HBV C Protein Phosphorylation in the Cell
Author:
Date:
2015-08-05
[Abstract]  Among the seven serines and one threonine in the carboxyl-terminus of HBV C protein, all but one (serine 183) appear in the context of RxxS/T consensus phosphoacceptor motifs and also overlap with other consensus motifs, such as S/TP, RS, SPRRR, RRRS/T, or RRxS/T, suggesting that various cellular kinases phosphorylate these residues. To determine whether threonine and/or serine (serines 157, 164, 170, 172, 178, and 180, and threonine 162, adw subtype) of HBV C protein are indeed phosphoacceptor residues in cells, Huh7 were transfected with a series of C-protein-expressing mutants, labeled with 32P-orthophosphate for 14 h, and then lysed. The 32Pi-labeled lysates were immunoprecipitated with anti-HBc antibody, and the 32Pi-labeled immunoprecipitated C ... [摘要]  在HBV C蛋白的羧基末端中的七个丝氨酸和一个苏氨酸中,除了一个(丝氨酸183)外,所有其它丝氨酸出现在RxxS/T共有磷酸受体基序的上下文中,并且还与其他共有基序重叠,例如S/TP,RS ,SPRRR,RRRS/T或RRxS/T,表明各种细胞激酶磷酸化这些残基。 为了确定HBV C蛋白的苏氨酸和/或丝氨酸(丝氨酸157,164,170,172,178和180以及苏氨酸162,adw亚型)是否确实是细胞中的磷酸受体残基,Huh7用一系列C- 蛋白表达突变体,用正磷酸32P标记14小时,然后裂解。 用抗HBc抗体免疫沉淀 32 P标记的裂解物,并通过放射自显影检测32 P标记的免疫沉淀的C蛋白。

Primer Extension Analysis of HBV DNA with Strand-Specific Primers
Author:
Date:
2015-08-05
[Abstract]  We performed primer extension assay to determine which steps of HBV DNA synthesis (i.e., minus- and plus-strand DNA synthesis and circularization of RC DNA) are affected by phosphoacceptor site mutations in C protein. In these experiments, we used several specific oligonucleotide primers. For quantitation, the level of extended DNA (ED) was normalized to the level of a single internal standard (IS) DNA. [摘要]  我们进行引物延伸测定以确定HBV DNA合成的哪些步骤(即,负链和正链DNA合成和RC DNA的环化)受C蛋白中的磷酸受体位点突变的影响。 在这些实验中,我们使用几种特异性寡核苷酸引物。 为了定量,将延伸DNA(ED)的水平标准化为单个内标(IS)DNA的水平。

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