Bioinformatics Analysis, Subcellular Localization and Toxicity Verification of Effector g11335 in Tilletia contraversa Kühn

doi:10.13560/j.cnki.biotech.bull.1985.2022-0414

Abstract

Abstract:

This research focused on analyzing the biological function of effector protein encoded by g11335 gene of Tilletia contraversa Kühn. Effector g11335 was an effector protein of GSDH family which may affect the plant healthy. The physicochemical properties, conserved domain, transmembrane domain, hydrophilicity and hydrophobicity of effector g11335 were analyzed using a variety of bioinformatics databases, and the expression vector g11335-pBin was constructed for subcellular localization analysis. The results showed that g11335 gene was 1 389 bp in length and encoded 462 amino acids. Subcellular localization of the effector protein showed that the protein was localized on the cell membrane. Tobacco toxicity test revealed that the effector protein was not toxic.

Key words: Tilletia contraversa Kühn, g11335, bioinformatics analysis, subcellular localization

GUO Zhi-hao, JIN Ze-xin, LIU Qi, GAO Li. Bioinformatics Analysis, Subcellular Localization and Toxicity Verification of Effector g11335 in Tilletia contraversa Kühn[J]. Biotechnology Bulletin, 2022, 38(8): 110-117.

Figures/Tables 10

Table 1 List of primers

引物名称 Primer name	引物序列 Primer sequence（5'-3'）
g11335-F	TCAGCACCAGCTAGCATCGATATGGCGCGCCAAC- TCGGCAAC
g11335-R	AACCGTTCATCGGCGGTCGACTCAAGCGGTTCCA- TCGATCGACA
g11335-YF	ATTTACGAACGATAGGGTACCATGGCGCGCCAAC- TCGGCAAC
g11335-YR	GCCCTTGCTCACCATGGATCCTCAAGCGGTTCCA- TCGATCGACA
pBin-F	ACAATCCCACTATCCTTCGC
pBin-R	AAGTCGTGCTGCTTCATGT
PGR107-F	ATAGCAGTCATTAGCACTTCCT
PGR107-R	CACTGGGGCATACTTCATGC

Fig. 1 Products of gene g11335 by PCR amplification M：DL2000 DNA maker

Fig. 2 Conserved domain analysis of effector g11335

Fig. 3 Hydrophilicity of effector g11335

Fig. 4 Signal peptide prediction of effector g11335

Fig. 5 Prediction of transmembrane region of effector g11335

Fig. 6 Secondary structure of effector g11335

Fig. 7 Tertiary structure prediction of effector g11335

Fig. 8 Subcellular localization of effector g11335 A: Empty carrier in the GFP channels. B: Empty carrier in the bright field of vision. C: Empty carrier in the compound field. D: g11335 in the GFP channels. E: g11335 in the bright field of vision. F: g11335 in the compound field（Scale: 20 μm）

Fig. 9 Toxicity validation of effector g11335

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