Identification and subcellular localization analysis of membrane protein Ycf 1 in the microsporidian Nosema bombycis

View article
Microbiology

Main article text

 

Introduction

Materials & Methods

Parasite and host

Cloning of Ycf 1 gene and analysis

Expression of recombinant protein of Ycf 1

Preparation of Ycf 1 monoclonal antibody and Western blot analysis

Immunolocalization of Ycf 1in N. bombycis

RNAi

Results

Cloning and expression of Ycf 1and immunoblot analysis

Specific detection of monoclonal antibody

Co-localization of Ycf 1 with SWP30 or Nb-actin in dormant spores of N. bombycis

Subcellular localization of Ycf 1 in proliferative phase of N. bombycis

Knockdown of Ycf 1 gene inhibited the proliferation of N. bombycis

Discussion

Conclusions

Supplemental Information

Dataset of standard curve line

DOI: 10.7717/peerj.13530/supp-1

Figure 6A and Figure S1 dataset

DOI: 10.7717/peerj.13530/supp-2

Figure 6B and Figure S2 dataset

DOI: 10.7717/peerj.13530/supp-3

Full length image of gels

DOI: 10.7717/peerj.13530/supp-4

Relative expression of Ycf 1 gene

(A) relative expression of Ycf 1 gene in RNAi group. (B) relative expression of Ycf 1 gene in control group. Error bars represent the standard deviations of three independent replicates. (One-way ANOVA analysis by 24h as control. n = 3, * p < 0.05, ** p < 0.01).

DOI: 10.7717/peerj.13530/supp-5

Copy number of Nbβ-tubulin after RNAi on Ycf 1.

(A) Copy number of Nb β-tubulin in RNAi group. (B) Copy number of Nb β-tubulin in control group. Error bars represent the standard deviations of three independent replicates. (One-way ANOVA analysis by 24h as control. n = 3, ** p < 0.01).

DOI: 10.7717/peerj.13530/supp-6

Statistical Reporting

DOI: 10.7717/peerj.13530/supp-7

Ycf 1 sequencing results

DOI: 10.7717/peerj.13530/supp-8

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests.

Author Contributions

Yong Chen conceived and designed the experiments, performed the experiments, authored or reviewed drafts of the article, and approved the final draft.

Erjun Wei performed the experiments, prepared figures and/or tables, and approved the final draft.

Ying Chen performed the experiments, prepared figures and/or tables, and approved the final draft.

Ping He performed the experiments, prepared figures and/or tables, and approved the final draft.

Runpeng Wang performed the experiments, prepared figures and/or tables, and approved the final draft.

Qiang Wang analyzed the data, prepared figures and/or tables, and approved the final draft.

Xudong Tang analyzed the data, prepared figures and/or tables, and approved the final draft.

Yiling Zhang analyzed the data, prepared figures and/or tables, and approved the final draft.

Feng Zhu analyzed the data, prepared figures and/or tables, and approved the final draft.

Zhongyuan Shen conceived and designed the experiments, authored or reviewed drafts of the article, and approved the final draft.

DNA Deposition

The following information was supplied regarding the deposition of DNA sequences:

The Ycf 1 sequences are available at NCBI GenBank: EOB13621.1.

https://www.ncbi.nlm.nih.gov/protein/EOB13621.1/

Data Availability

The following information was supplied regarding data availability:

The raw data are available in the Supplemental Files.

Funding

This work was funded by the China Agriculture Research System of MOF and MARA and the National Natural Science Foundation of China (31960684). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

3 Citations 1,069 Views 103 Downloads

Your institution may have Open Access funds available for qualifying authors. See if you qualify

Publish for free

Comment on Articles or Preprints and we'll waive your author fee
Learn more

Five new journals in Chemistry

Free to publish • Peer-reviewed • From PeerJ
Find out more