Protein coexpression using FMDV 2A: effect of "linker" residues.
Protein coexpression using FMDV 2A: effect of "linker" residues.
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DOI：
10.1155/2013/291730
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发表时间：
2013
影响因子：
--
通讯作者：
Ryan MD
中科院分区：
文献类型：
--
作者：
Minskaia E;Ryan MD
Many biomedical applications absolutely require, or are substantially enhanced by, coexpression of multiple proteins from a single vector. Foot-and-mouth disease virus 2A (F2A) and “2A-like” sequences (e.g., Thosea asigna virus 2A; T2A) are used widely for this purpose since multiple proteins can be coexpressed by linking open reading frames (ORFs) to form a single cistron. The activity of F2A “cleavage” may, however, be compromised by both the use of shorter versions of F2A and the sequences (derived from multiple-purpose cloning sites) used to link F2A to the upstream protein. To characterise these effects, different lengths of F2A and T2A were inserted between green and cherry fluorescent proteins. Mutations were introduced in the linker region immediately upstream of both F2A- and T2A-based constructs and activities determined using both cell-free translation systems and transfected cells. In shorter versions of F2A, activity may be affected by both the C-terminal sequence of the protein upstream and, equally strikingly, the residues immediately upstream introduced during cloning. Mutations significantly improved activity for shorter versions of F2A but could decrease activity in the case of T2A. These data will aid the design of cloning strategies for the co-expression of multiple proteins in biomedical/biotechnological applications.
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影响因子：
3.7
作者：
Kim JH;Lee SR;Li LH;Park HJ;Park JH;Lee KY;Kim MK;Shin BA;Choi SY
通讯作者：
Choi SY
DOI：
10.1073/pnas.0710650105
发表时间：
2008-03-25
影响因子：
11.1
作者：
Baranick, Brian T.;Lemp, Nathan A.;Logg, Christopher R.
通讯作者：
Logg, Christopher R.
影响因子：
20.3
作者：
Huang, X;Wilber, AC;Zhou, XZ
通讯作者：
Zhou, XZ
影响因子：
4.5
作者：
de Felipe, P;Ryan, MD
通讯作者：
Ryan, MD
影响因子：
--
作者：
Pfuetzner, Wolfgang
通讯作者：
Pfuetzner, Wolfgang