ligation independent cloning-分子克隆技术ppt课件 LIGATION-INDEPENDENTCLONING 1 经典的克隆方法 经典的克隆方法，主要分为以下两种：(1)粘性末端连接，即载体质粒和待插入的外源片段都通过合适的限制性内切酶切割，产生相互匹配的粘性末端，然后在连接酶作用下重新形成磷酸二酯键而得到环化的重组质粒；(2)平末端连接...
They also show a high degree of homology (40–60% identical residues) with their counterparts from other insect vectors, such as Anopheles gambiae and Aedes aegypti. The mRNA expression profiles of these six proteases vary considerably: two trypsin-like proteases (Pptryp1 and Pptryp2) are ...
(DOI: 10.5376/.2011.30.0013) ligation-independent Vectors Vector Tag Expression Host Resistance Tag(s) Size 2BT His6-tev-yORF E.coli T7 AMP 2 kDa 2CT His6-MBP-N10-tev-yORF E.coli T7 AMP 45 kDa 2GT His6-GST-tev-yORF E.coli T7 AMP 28 kDa 2NT His6-NusA-tev-yORF E.coli...
方式不仅克服了常规依赖连接酶克隆方法的缺点 而且操作简便 方法快捷及连接效率高 应用日益广泛 在T4DNA聚合酶的3 5 外切酶活性下和一种特定的dNTP存在的缓冲液条件下反应 插入片段能产生10 15个碱基的粘性末端 AslanidisanddeJong 1990 载体和PCR片段依靠长的粘性末端重组 此过程不需要连接酶 这种克隆方法不需要考虑...
Transient transfections of Rat2 cells with transcription factor expression vectors allowed identifying KLF15 as a putative repressor of T1 transcript in Rat2 cells. Conclusion Altogether, these data contribute to a better knowledge of fut8 expression in YB2/0 that will be useful to better control ...
Several other laboratories have deposited pLKO derived vectors that may also be useful for your experiment. To see these vectors, visit Addgene’s website and “search for “pLKO”“. Back to Top B. Designing shRNA Oligos for pLKO.1 ...
ligationindependentcloning-分子克隆技术ppt课件 1 经典的克隆方法经典的克隆方法 经典的克隆方法,主要分为以下两 种:(1)粘性末端连接,即载体质粒 和待插入的外源片段都通过合适的 限制性内切酶切割,产生相互匹配 的粘性末端,然后在连接酶作用下 重新形成磷酸二酯键而得到环化的 重组质粒;(2)平末端连接,平末端 连接...
Central polypurine tract, cPPT, improves transduction efficiency by facilitating nuclear import of the the transduced cells. Human phosphoglycerate kinase promoter drives expression of puromycin. Puromycin resistance gene for selection of pLKO.1 plasmid in mammalian cells. 3’ Self-inactivating long ...
For example, some vectors have high copy numbers and will produce large amounts of subcloned DNA inserts. Others have been designed to facilitate in-vitro transcription and super-expression of proteins in-vivo. Subcloning involves the ligation of a previously cloned and purified DNA molecule into...
Mutagenesis of the wo CRE and/or cotransfection with vectors that express ICERII'Y or CREM'T indicated that the two CRE have major roles in 'PT-I expression. The two CRE are also required for optimal promoter activity by SCF and IL-Ia. A particular cytokine could oncomitantly induce ...