Molecular Characterization and Expression Analysis of Four Janus Kinases (JAK1, JAK2a, JAK3 and TYK2) from Golden Pompano (Trachinotus ovatus)
Round 1
Reviewer 1 Report (New Reviewer)
This manuscript (fishes-2352575) reports the full-length cDNA sequences of JAK1, JAK2a, JAK3 and TYK2 from golden pompano and their gene expression analysis. This study could contribute as a first step toward understanding the immune system of this fish species. I have several minor comments on the publication of this paper, which are as follows.
1. Since JAKs are known to be involved in the regulation of inflammatory and antiviral cytokines as signaling molecules of the immune system in mammals, I recommend that inflammatory and antiviral cytokines also be examined in gene expression analysis of LPS (inflammatory cytokines) and Poly I:C (antiviral cytokines) stimulation and V. alginolyticus infection (inflammatory cytokines), together with JAKs. This will help us to better understand the role of JAKs in the immune system of this fish species.
2. Although this study is limited to gene structure, phylogenetic tree analysis and gene expression analysis of JAKs, I suggest that the authors describe in a discussion or conclusion what future research is needed to elucidate the role of JAKs in the immune system.
3. LPS and Poly I:C are immune stimulants, not pathogens, so "infection" in the text should be rephrased as "stimulation".
English language fine, However, I suggest that LPS and Poly I:C are immune stimulants, not pathogens, so "infection" in the text should be rephrased as "stimulation".
Author Response
Dear Reviewer:
Thanks a lot for your laborious work on the paper titled “Molecular characterization and expression analysis of four Janus kinases (JAK1, JAK2a, JAK3 and TYK2) from golden pompano (Trachinotus ovatus)”, which make the paper’s quality increased greatly.
We are so grateful for that and we have carefully revised the paper according to the comments.
Please see the attached file.
Author Response File: 
Author Response.pdf
Reviewer 2 Report (New Reviewer)
Janus kinases play important roles in immune responses of vertebrates, though their role in diverse fishes is yet being investigated. Xie et al. sequenced and characterized four JAK genes in golden pompano and characterized their expression in multiple tissues following challenge with different antigens. Their results show that the respective genes are differentially involved in immune response of the species. This is a straightforward – though technically challenging – study, and the results are defensible. I have context-specific comments below. I’ve marked the manuscript to guide revision of the prose.
Abstract. – At line 17, why are the authors writing of pharmacological mechanisms? This may be a poor translation from Chinese. Pharmacological mechanism is the process by which a drug works in the host. Perhaps these few words can simply be removed without loss of meaning.
I suggest that the last sentence of the abstract read: Knowledge of the roles of JAKs in the immune response to different microbial pathogens provides a basis for further understanding of these functions.
Introduction. – I have marks for strengthening the prose on the manuscript document.
Methods. – The URL for DNAstar software should be added at line 90 and can be deleted from line 102.
A supporting citation or URL should be provided for MEGA at line 106.
At line 132, “all assumptions” is quite vague, and something more substantive should be added.
Results. – The captions for the figures should be in present tense – these results are being presented as the reader sees them.
At line 200, the Latin name for Asian seabass should be given.
Figure 5 is just too small to read and should be presented as a larger image.
Discussion. – I wrote some editorial marks on the manuscript document.
References. – I noted a small number of issues with the citations on the manuscript document.
Comments for author File: Comments.zip
I’ve marked the manuscript to guide revision of the prose. Like many Chinese speakers, the authors are confused about use of articles ("a" and "the") and past and present tense. There's awkward wording in many contexts.
Author Response
Dear Reviewer:
Thanks a lot for your laborious work on the paper titled “Molecular characterization and expression analysis of four Janus kinases (JAK1, JAK2a, JAK3 and TYK2) from golden pompano (Trachinotus ovatus)”, which make the paper’s quality increased greatly.
We are so grateful for that and we have carefully revised the paper according to the comments.
Please see the attachment..
Author Response File: Author Response.pdf
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
Round 1
Reviewer 1 Report
The paper is generally well written and the main idea is to explain the role of fish JAKs due to a challenge against different pathogens. The structure is correct, making it easy to read and understand. Bibliographic references are current and adequate.
I found some mispellings and a suggestions. Please review the attach.
Comments for author File: Comments.pdf
Author Response
Dear Reviewer,
Thanks a lot for your laborious work on the paper titled “Molecular cloning and expression analysis of four Janus kinases (JAK1, JAK2a, JAK3 and TYK2) from golden pompano (Trachinotus ovatus)”, which make the paper’s quality increased greatly.
We are so grateful for that and we have carefully revised the paper according to your comments.
Reviewer #1: The paper is generally well written and the main idea is to explain the role of fish JAKs due to a challenge against different pathogens. The structure is correct, making it easy to read and understand. Bibliographic references are current and adequate.
I found some mispellings and suggestions. Please review the attach.
Line 61：In vivo should be in italics.
Response: It had been revised.
Line97: You should indicate LPS strain used.
Response: It had been revised as suggestion.
Table 1: Suggestion: Table was divided into techniques used. I mean, you should prepare one table to nested-PCR, other to RACE-PCR and another to qPCR. In this way, the table should be adjustment of the size contents.
Response: It had been revised as suggestion.
Figure 2. Suggestion: All the graphs should be at the same scale, so maybe you should represent (0-15) in the Y axis.
Response: It had been revised as suggestion.
Figure 3. Suggestion: All the graphs for the same gene at least, should be at the same scale, so maybe you should represent (0-10) in the Y axis.
Response: It had been revised as suggestion.
Line 325: This bibliography is not correctly.
Response: It had been revised.
Line 414: This bibliography is not correctly.
Response: It had been revised.
Best wishes,
Reviewer 2 Report
According to my revision, the manuscript deserved publication as it has been submitted. No relevant comments on it.
just a minor suggestion: change figures from with and black to colour, this will increase the visual quality of the ms
Author Response
Dear Reviewer,
Thanks a lot for your laborious work on the paper titled “Molecular cloning and expression analysis of four Janus kinases (JAK1, JAK2a, JAK3 and TYK2) from golden pompano (Trachinotus ovatus)”, which make the paper’s quality increased greatly.
We are so grateful for that and we have carefully revised the paper according to your suggestions.
Reviewer #2: According to my revision, the manuscript deserved publication as it has been submitted. No relevant comments on it.
just a minor suggestion: change figures from with and black to colour, this will increase the visual quality of the ms
Response: It had been revised as suggestion.
Best wishes,
Reviewer 3 Report
1. Why did the authors choose this technology instead of other GWAS? Please provide the reason.
2. What cell type was utilized for RNA extraction
3. How much RNA was used in PCR? Was it one-step or 2-step PCR?
4. The abstract is not particularly informative and would benefit from more background.
5. Summarize the abstract, focus on the main findings and mention the small conclusion in at the end of abstract
6. In the Introduction focus on the objectives and insert a few new reference and relevant findings
Author Response
Dear Reviewer,
Thanks a lot for your laborious work on the paper titled “Molecular cloning and expression analysis of four Janus kinases (JAK1, JAK2a, JAK3 and TYK2) from golden pompano (Trachinotus ovatus)”, which make the paper’s quality increased greatly.
We are so grateful for that and we have carefully revised the paper according to your comments and suggestions.
Reviewer #3: 1. Why did the authors choose this technology instead of other GWAS? Please provide the reason.
Response: The reasons accounting for choosing this technology described in the manuscript are as follows：
Firstly, the cost of this technology is relatively low compared to other technologies.
Secondly, we consider this technology is easier to perform than other technologies.
Last but not least, our lab is more familiar with this technology than any other.
- What cell type was utilized for RNA extraction
Response: Spleen tissues. It had been revised in section 2.3.
- How much RNA was used in PCR? Was it one-step or 2-step PCR?
Response: The RNA used in PCR was 1.19 μg and the methods of one-step or 2-step PCR had been described in detail in section 2.3.
- The abstract is not particularly informative and would benefit from more background.
Response: It had been revised as suggestion.
- Summarize the abstract, focus on the main findings and mention the small conclusion in at the end of abstract
Response: It had been revised as suggestion.
- In the Introduction focus on the objectives and insert a few new reference and relevant findings
Response: It had been revised as suggestion.
Best wishes
Round 2
Reviewer 3 Report
i am sorry to say i can,t Recommended your paper to post in this journal
