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Characterization of a Pinus sylvestris thaumatin-like protein gene and determination of antimicrobial activity of the in vitro expressed protein

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Abstract

Thaumatin-like proteins (TLPs) are pathogenesis-related proteins, which are involved in plant defense responses to pathogen infection. Expression of the Pinus sylvestris L. TLP gene is up-regulated by methyl jasmonate treatment and inoculation with Heterobasidion annosum. A full-length Pinus taeda TLP gene sequence was used to design PCR primers for amplification of the full-length TLP gene from P. sylvestris. A putative 705-bp open reading frame of TLP gene was cloned into Escherichia coli cells, and then subcloned into the overexpression vector pET100 using BL21 Star expression bacteria. Optimization of the expression of recombinant TLP was achieved by decreasing both expression temperature and IPTG concentration. The purified 24.6-kDa TLP shows antimicrobial activity against 12 fungal species.

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Funding

This work was funded by Latvian Council of Science project “Investigation of molecular defence mechanisms in Scots pine (Pinus sylvestris L.)” (No. 284/2012).

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  1. Latvian State Forest Research Institute “Silava”, Rigas 111, Salaspils, LV-2169, Latvia

    Ilze Šņepste, Vilnis Šķipars, Baiba Krivmane, Lauma Brūna & Dainis Ruņģis

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  1. Ilze Šņepste
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  2. Vilnis Šķipars
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  3. Baiba Krivmane
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  5. Dainis Ruņģis
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Correspondence to Dainis Ruņģis.

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Communicated by P. Ingvarsson

Data archiving statement

The DNA sequence of the P. sylvestris TLP gene has been deposited in GenBank (Acc. No. JX461338).

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Šņepste, I., Šķipars, V., Krivmane, B. et al. Characterization of a Pinus sylvestris thaumatin-like protein gene and determination of antimicrobial activity of the in vitro expressed protein. Tree Genetics & Genomes 14, 58 (2018). https://doi.org/10.1007/s11295-018-1274-9

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