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Expression of the small cysteine-rich protein SCR96 from Phytophthora cactorum in mammalian cells: phytotoxicity and exploitation of its polyclonal antibody

Biotechnol Lett. 2020; 
Huang SX,, Zhang ZH,, Liu W,, Tao H,, Zhang Y,, Shi NX,, Zhu F,, Ji ZL,, Chen XR,.
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Gene Synthesis … Then the gene fragment containing EcoRI and HindIII on the 5′- and 3′-end respectively (for the DNA sequence see Supplementary Fig 1) was synthesized by Genscript and cloned into pUC57-Simple … No M0101, GenScript) as a positive control … Get A Quote

摘要

We aimed to investigate the expression of a novel small cysteine-rich (SCR) effector protein SCR96 from the phytopathogenic oomycete Phytophthora cactorum in mammalian cells, its bioactivity and to exploit its polyclonal antibody.,The gene encoding the SCR effector protein SCR96 was codon-optimized, custom-synthesized, cloned into pcDNA3.1(-) and overexpressed in human embryonic kidney (HEK) 293-6E cells. The recombinant protein SCR96 was prone to aggregation and purified with its monomer to homogeneity with a predicted molecular weight of 8.9 kDa. SCR96 exhibited strong phytotoxic activity on tomato seedlings at 24 h post treatment with 4.2 μg of the purified protein. An anti-SCR96 polyclonal antibody was ... More

关键词

Ectopic expression; Mammalian cell; Phytophthora cactorum; Phytotoxicity; Polyclonal antibody; Small cysteine-rich protein