The PBC Program for Fellowships for Outstanding Post-doctoral Fellows

from China and India - 2014/2015

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Application Form

INSTRUCTIONS:
The application form and all supporting documents must be typed in English and submitted to the
university, according to the guidelines determined at each institution, together with a recent
photograph. The application must be signed by the candidate. In addition, the following supporting
documents, typed in English, are required:
1. Curriculum vitae including a list of publications (Please note the impact factor of each
publication, if exists).
2. Three letters of recommendation typed in English from three referees, at the request of the
candidate.
3. Commitment of a senior faculty member from the designated Israeli university indicating
his/her agreement to guide the candidate.
4. All relevant diplomas (bachelor, master and PhD).

A) PERSONAL INFORMATION
1. Full Name

2. Citizenship

3. Passport number

8818849

4. Date of birth
5. Place of birth
6. Permanent address

1/

home:

7. Phone numbers

work:
mobile:
milton.sxc@gmail.com

8. E-mail address
9. Estimated date of arrival in Israel
10. Gender
Male
11. Marital Status (if married, include
name of spouse)
12. Number and age of children

13. Which

family

members

will

accompany you?
14. Present appointment and place of Post Doctoral Student at Huazhong Agricultural
employment

University, Wuhan, China

15. Knowledge of languages (none, low,

fair, high)

B) ACADEMIC CAREER
16. Academic discipline
17. Academic
degrees

studies

and

Language Reading
English

Hebrew
Other
Tamil

Writing

Speaking

university Ph.D: Manonmaniam Sundaranar University,

(please

indicate

were

conferred

the Tirunelveli, Tamilnadu

institution).
If

degrees

honours, please indicate.

18. Title of PhD dissertation

with
Aquaculture

Potentials

of

Threatened

Snakehead by Seed Production and Larvi

19. Submission

of

PhD

Culture Technologies
thesis

(month/year)
20. PhD graduation (month/year)

If not yet approved, please provide

anticipated date.
21. Academic awards and distinctions

CSIR Senior Research Fellowship

C) PLANNED

STUDY

AND

RESEARCH
22. Preferred department to conduct your Dr. Dror Angel,
studies/research

The Leon Recanati Institute for Maritime

23. Period of proposed research or study

Studies

D) FINANCE
24. Do you expect to receive a salary,
partial salary, scholarship, fellowship, Expecting the Stipend / Salary USD2500 /
grant, sabbatical allowance, study month
allowance or any other payment from
any source, including your present
university or any other source during
the proposed period? If so, please
provide

details

of

any

such

allowances.
25. You are highly encouraged to apply
for a fellowship, scholarship or grant
from

another

institution.

Please

indicate to which fellowships, if any,

you have applied.
Please Note: The financial aid of this scholarship is adequate only for the living expenses
of the fellow himself/herself.

26. Please provide a summary of your dissertation (up to 2 pages).

Title of thesis
Seed

: Aquaculture Potentials of Threatened Snakehead by

Production and Larvi Culture Technologies

Name of the Candidate

: M.James Milton

Registration Number

: 2712

Name of the Supervisor

: Dr .M.A.Haniffa
CSIR Emeritus Scientist & Director
Centre for Aquaculture Research and Extension (CARE)
St.Xaviers College (Autonomous), Palayamkottai 627
002.

Abstract
Murrels belonging to the family Channidae, vary greatly in size at maturity. They
are economically important, occupying the top rank in South East Asia due to their taste,
medicinal qualities and less intramuscular spines. So far, no attempt on induced breeding
has been made on threatened murrel C. gachua and the present investigation is the first of
its kind dealing with seed production and larviculture of a threatened murrel species
Channa gachua. Morphometric and meristic characters of three different populations of C.
gachua were analysed for the cluster analysis. Tamirabarani and Godavari populations were
found to be closer when compared to Brahmaputra population. This may be due to similar
environmental condition and water bodies being in closer association. Brahmaputra river
has a different origin with altogether different environmental conditions, with more rainfall.
The gonadosomatic indices of females varied significantly between different months for
Tamirabarani River populations. The GSI value declined rapidly from 3.31 to 0.79 after
spawning. Therefore it was confirmed that the fish spawned once in a year with spawning
peak during December to February. However, in the present study, six oocytic stages (I to
VI stages) were clearly defined. It can also be inferred from the observed findings that in
Channa gachua there occurred atleast 3 phases in the annual ovarian activity. In the present
study, three different types of hormones viz: HCG, LHRHa and Ovaprim with different
doses (Low, Medium and High) were used for induced spawning. Considering the overall
performance of the hormones with regard to number of eggs spawned (2471 265.82),
latency period (21.3 0.18 h), fertilization rate (70%) and hatching rate (72%), HCG was
found to be the most potent ovulating agent in C.gachua. The early post larvae of C.
gachua were fed with four different types of feed with and without fish meal for 45 days at

a stocking rate of 20 individuals in a glass tanks. At the end of 45 th day, growth rate was the
best and significantly higher in fishes fed on Chicken Intestine mixed diet (0.064mg/g/day)
followed by Control (0.053mg/g/day) whereas a low growth rate was observed in fish fed
on Jawala (0.048mg/g/day). The analyses of the muscle tissues of C. gachua revealed the
presence of twelve amino acids. Hence it is suggested that chicken intestine is a better
supplementary material than fish meal since it has produced better growth in C. gachua
comparing with anchovy and jawala diet. The present finding suggests that the threatened
murrel C .gachua could be conserved through induced breeding techniques.
27. Please provide a summary of your present research (if different from your dissertation).
Isolation of RNA from healthy and diseased freshwater fishes
using Trizol reagent.
cDNA library construction using GS FLX technology
Immune gene identification from the cDNA library through the BLAST2GO program.
Bioinformatic analysis of the identified immune genes using various tools.
The relative expression of Immune in blood, gills, liver, heart, spleen, intestine, head
kidney, kidney, skin, muscle and brain were measured by quantitative real time
polymerase chain reaction
Cloning of Immune gene
Over expression and purification of immune protein.
28. Please provide a summary of your research plan (up to 4 pages).
TITLE : Construction Of Cdna Library And Molecular Characterization Of Immune
Genes From Litopeneaus vannamei Against White Spot Disease .
1

Objectives and Significance of the Proposed Research

Objectives:
1.
2.
3.
4.
5.
6.
7.

To construct cDNA library of healthy and infected Peneaus vannamei

To compare the transcriptome data of normal and healthy shrimp.
To identify the immune genes from the constructed cDNA library.
To analyse the immune gene using bioinformatic tools.
To clone the identified immune genes.
To purify and analyse the activity of the recombinant protein.
To find out the expression of the identified genes from different tissues.

Methods
Isolation of Bacteria and Virus:
Total RNA extraction

Different tissues (Liver, Kidney, Spleen, Intestine, Heart, Gills, Brain, Skin, Muscle and
Blood) will be collected from both normal and Viral infected Peneaus vannamei. Total
RNA will be extracted from each tissue using one step RNA reagent method.
cDNA library construction
Total RNA will be used for cDNA synthesis from
i)

Normal Peneaus vannamei,

ii)

Viral infected Peneaus vannamei.

cDNA library will be constructed using pyrosequencing technology. The cDNA library will
be annotated using BLAST2GO and transcriptome datasets will be constructed.

Comparative study of transcriptome datasets

The transcriptome dataset of the infected fish will be compared with that of the normal fish
to identify the up regulated and down regulated genes.
Bioinformatics analysis
The Basic Local Alignment Tool (BLAST) program will be used to search similar
nucleotide and protein sequences. The open reading frame (ORF) and amino acid sequence
will be obtained by using DNAssit 2.2. Characteristic domains or motifs would be
identified using the PROSITE profile database. Identity, similarity and gap percentages
would be calculated using FASTA program. The N-terminal transmembrane sequence will
be

determined

by

DAS

transmembrane

prediction

program

(http://

www.sbc.su.se/wmiklos/DAS). Signal peptide analysis will be done using the SignalP

worldwide P server (http://www.cbs.dtu.dk). Pairwise and multiple sequence alignment
would be analyzed using the ClustalW version 2 program. The phylogenetic relationship of
the Immune Gene will be determined using the neighbor-joining (NJ) method and MEGA
4.1 program. The three dimensional structure of the depicted protein will be predicted using
I Tasser and analysed using PyMol.
Submission of gene in EMBL
The identified immune genes of Peneaus vannamei will be submitted to EMBL database.
Cloning of identified virulence genes
The identified immune genes will be amplified by gene specific primers using thermal
cycler. The amplified product will be cloned into an appropriate bacterial vector (pET28a,

pRSETa etc) (Sambrook et al., 1989). The positive clones will be sequenced to ensure
inframe insertion. The cloned plasmid will be transformed into bacterial host (E.coli) for
over-expression.
Over expression of cloned genes
For large scale production of recombinant protein, the transformed clones will be subjected
to induction with the respective inducer (E.g., IPTG or Arabinose) for overexpression
(Miroux et al., 1996).
Protein purification
The vectors that will be employed in cloning will possess a fusion tag (E.g., His-tag, GSTtag, MalE-tag), which will also be expressed along with the protein. These fusion tags will
bind to their respective ligands present in the matrix. Thus, the protein expressed by over
expression will be purified through affinity chromatography.
Protein assays
Protein specific assays will be performed to confirm the immune nature of the recombinant
immune proteins (Kurz et al., 2003).

Time schedule of Research activities

Period of study
1 2 Months

Achievable targets
Isolation of Aeromonas pathogen and infection of Misgurnus

2 - 6 Months

anguillicaudatus
cDNA libraries Construction

6 - 12 Months

Immune genes identification & Bioinformatic Characterization

12 - 16 Months

Cloning of immune genes

16 - 20 Months

Protein expression

20 - 24 Months

Recombinant Protein characterization

24 - 26 Months

Report Submission

29. Please explain, in some detail, the reasons for your choice of institution and academic
supervisor and the connection, if any, with your present and planned future work (10-12
lines).
Dr. Dror Angel is doing excellent research on Aquaculture. His specialisation is very helpful for
my research work. Under his guidance I will do excellent research work on shrimp. This
research work may very much helpful of the farmers.

30. Please describe in one paragraph (10-12 lines) what you hope to have achieved at the
conclusion of your post-doctoral work.
Research achievements (Germplasm, genes, knowledge etc)
-

cDNA library will be constructed.

Immune-related genes will be identified.
Immune-related genes expression level will be measured.
Clones with immune genes.
Recombinant immune proteins.

I will Publish Atleast 3 articles in SCI journals.

I will try file patent also.
I have read the rules and guidelines in their entirety, confirm that I shall abide by them and agree
to provide any further information which the PBC steering committee may deem necessary to
evaluate my candidacy.
26/10/2013
Date

M. James Milton
Signature