The cDNAs of EGFPpHΔC12 and NlucΔN4 were amplified by PCR and fused with the intermediate random linker sequence of two amino acids using Hot Fusion methods (Fu et al. 2014). The linker sequence “W-T” was selected using an in-house random screening system. The cDNA fragments encoding...
Our results show that active IBs of PhoC can be generated by GFP fusion, demonstrating for the first time the potential of GFP fusion to induce active IB formation of another soluble protein. We also show that the linker sequence in PhoC-GFP fusion proteins plays an important role on the ...
and 3D structures of the minimal Drp1 dimer (PDB ID: 4BEJ) and mEGFP (PDB ID: 2Y0G) drawn to scale. A Drp1 monomer is color-coded in correspondence with the above domain arrangement. The sequences of the linkers connecting mEGFP to Drp1 at the N- and C...
Recombinant Fusion Proteinsp pBackground/p pInteins are proteins that catalyze their own removal from within larger precursor proteins. In the process they splice the flanking protein sequences, termed the N-and C-terminal exteins. Large inteins frequently have a homing endonuclease that is ...
8b). The half-life of GPI-anchored proteins is mainly modulated by the phospholipase family of proteins28, and a recent study described that a protein domain adjacent to a GPI-attachment signal sequence defined the activity of phospholipases on GPI-anchored proteins29. Thus, mutated protein ...
Yeast DHODH and Aequorea GFPmut2 (1) coding sequences were inserted into anE. coli-Plasmodium falciparumshuttle plasmid (pHHMC*/3R0.5) (2), such that DHODH-GFP were fused into a single coding sequence (including a 10Ala-1Gly linker segment). ...
We successfully obtained intein-mediated purified GFP1–9 (ipGFP1–9) with improved expression levels and high solubility by designing the linker sequence. We then investigated and characterized ipGFP1–9 for reconstitution with S10 and S11, enabling in vitro reconstitution applications using ...
然后通过融合 PCR 技术将 Gluc 基因与 G11 基因通过 Linker 连接并插入到 pcDNA3.1 上,与 G1-10 表达质 粒共转染 293T 细胞,测定细胞上清中 Gluc 荧光素酶的活性,并进行 GFP 荧光 观察。为了更方便对病毒进行细胞水平的观测和筛选,利用慢病毒表达系统构建 能稳定表达 G1-10 的 MDCK 细胞系,再向细胞系中...
(1) DNA sequences coding for test protein X is randomly mutated and recombined by DNA shuffling. (2) The library of mutants is cloned into the GFPcp reporter plasmid, transformed into E. coli, and the colonies induced with IPTG to express the X-GFPcp fusion protein. (3) Brighter clones...
the gfp gene cassette was inserted into a gene homolog ofS. typhimuriumATP-dependent helicase protein (hrpA). In the fluorescentSalmonella typhimurium, the gfp gene cassette was inserted into the sequence of a gene encoding a common antigen found in the outer membrane ofSalmonellaand other enterobac...