Centre of Excellence in Molecular Biology

University of the Punjab, Lahore-Pakistan

CERTIFICATE

It is certified that Yousuf Shafiq, Reg. No. 2014-ag-5019, student of B.Sc (Hons.) Agri. Sciences,
(Plant Breeding and Genetics) University of Agriculture, Faisalabad has completed his internship of 8
weeks (from 19 Feb to April 23, 2017) under my supervision at Centre of Excellence in Molecular
Biology, University of the Punjab, Lahore-Pakistan. He has completed all the work necessary at this
level.

He is hardworking and technically sound and result oriented. He performed his tasks with great
responsibility and showed excellent performance. He is very enthusiastic and willing to learn. I wish
him best of luck for his future endeavors.

SUPERVISOR AT CEMB: _____________________

Dr. Idrees Ahmad Nasir
Associate Professor
Seed Biotechnology Lab

_______________________
COORDINATER:
Dr. Mudasir Fareed
Ph.D. Scholar

INTERNAL SUPERVISOR: _______________________

Dr. Tanvir Malik
Professor
Department of PBG, UAF

CHAIRMAN: _______________________
Prof. Dr. Hafeez Sadaqat
Department of PBG, UAF
 1. CEMB Profile:

On December 09, 1981, Chairman University Grants Commission (UGC) convened a meeting
of fifteen eminent professors to discuss ways and means to promote teaching and research in
the emerging science of molecular biology. The members unanimously recommended
establishing an autonomous institution of advanced teaching and research on the campus of
University of the Punjab, Lahore. In November 1985 the proposal to establish the Centre for
Advanced Molecular
Biology was approved at a cost of 24.55 million rupees. In 1986, the project was upgraded
into a Centre of Excellence in Molecular Biology (CEMB) and the cost of setting it up was
subsequently revised in January 1991, to 44.33 million rupees. In April 1987, the Federal
Ministry of Science Technology (MOST) approved the establishment of a Centre for Applied
Molecular Biology (CAMB), located back to back with the laboratory block of the Centre of
Excellence in Molecular Biology (CEMB).

The CEMB established nucleus laboratories in March, 1985 in two student laboratories of the
Department of Zoology, University of the Punjab, Quaid-e-Azam Campus, Lahore.
Construction work on the CEMB building started in 1987 on a site located in Lahore's southern
suburb of Thokar Niaz Baig along the Canal. The scientific staff moved into the new building
in late 19921993.
The CEMB laboratory complex is spread over 60 acres of land, with a covered area of 115,000
square feet, including an old Laboratory Block, four new Laboratory Blocks, a Teaching
Block, and Hostels for Local Foreign Research Scholars. The Laboratory Blocks comprising
a total of 70 Research Labs, 16 Containment Labs, one Production Unit and Support Facilities,
comprising a Lab-Aid Section (for washing, autoclaving and media preparation), Repair and
Maintenance Section, Engineering Section, an Animal House, an Insectary, six large Plant
Growth Rooms, and two stories store for research materials. The Teaching Block consists of
a well-equipped Library cum Reading Room, three Seminar Lecture Halls, and five
Committee Rooms, one Library cum Reading Room, one Computer Lab and Modern Facilities
for Photocopy Photography. Administration comprises of Director's Office, Visitors Room
and Administration Office and Accounts Office. There is a self- service Canteen, two Dining
Halls and 55 rooms each for two students in the research scholar’s hostels.

2. Functional Setup:
 3. Objectives:
The Centre has addressed itself to the following functions:
 Teaching and training to generate a cadre of manpower specifically trained in
molecular biology and recombinant DNA technology.
 Undertake goal oriented molecular biological research on specific problems, in
agriculture, health & medicine, industry, energy and environmental sectors.
 Create a repository of DNA enzymes, DNA cloning vectors, novel bacterial strains and
other such molecular tools for ready availability and use by various research groups at
this Centre and other DNA research laboratories in Pakistan.
 Organize national and international seminars and conferences for in-depth discussions
on scientific and technological developments which will lead to new ideas and
 innovative applications of knowledge in gene cloning and recombinant DNA
technology.

4. Achievements:

Produced 206 M.Phils. and 40 Ph.Ds. with present enrollment at 134. More than 500 scientists
from Pakistan and member countries of OIC & SARC (Bangladesh, Egypt, India, Iran, Jordan,
Nepal, Sri-Lanka and Turkey) have passed through the corridors of the Centre to learn various
methodologies and/ or spend sabbatical assignments. Thus the Centre has assumed the role of
a regional training Centre.

 Developed Bt transgenic rice that is in the “field trials”.

 Developed Bt cotton that is in the “field trials”.
 Developed Bt pesticide formulation.
 Synthesized four Bt pesticidal genes with optimized codon usage and developed plant
expression vectors containing 2-3 genes.
 Obtained 5 patents and 3 patents are in the process.
 Established techniques which are being used to raise disease free seeds of gladiolus,
sugarcane, potato and even fungus free stock of gladiolus.
 Cloned human pharmaceutical protein genes for commercial production.
 PCR based diagnosis of tuberculosis, hepatitis, SARS, HIV, S. typhi, H. Pylori etc
available to the general public as economical and reliable diagnostic procedures.
 Discovered 11 new deafness loci, 3 new vision impairment loci, 17 new genes and one
novel deafness modifier gene.
 Developed procedures for;
• The prenatal diagnosis of b-thalassemia.
• DNA Typing for human identification that are being used in crime investigation
and parenthood confirmation. [Processed 334 cases of parentage, rape, murder,
dead body identification and robbery].
 5. Research Work on Variety Development:
Most of the research work at CEMB is of molecular biology. In plant molecular biotechnology
section, breeding work is very less. It has been initiated seriously some 4-5 years ago. Main
obstacle in breeding was the unavailability of a breeder. For solving this problem, Dr. Ghulam
Abbas has been recruited as breeder. Now the breeding work is on three main crops, cotton,
maize, tomato and sugarcane. Transgenic cotton having Bt gene is being crossed with other
standard varieties to develop a variety that could be insect resistant and good yield with quality
parameters at the same time.

The cross is handled through conventional breeding techniques. For example, a recent line of
Bt cotton CEMB-33 has been developed by crossing CEMB-02 with N-1148. This variety has
been approved recently in December 2013. CEMB also has collaboration with a multinational
company Ali Akbar Group for variety development and seed multiplication. The center
provides the pre-basic seed to this company for multiplication and seed distribution. At present
research is also going on double Bt and triple Bt cotton with an aim to increase the expression
level of Bt gene in these varieties. Similarly research is being conducted on transgenic hybrid
maize and tomato and insect pest resistant sugarcane.

6. Research Wings:

There are three main research wings at CEMB that are further divided into various sections

a. Plant Molecular Biology o Transgenic plant technology

o Quality seed production o Search for drought tolerant
genes o Microarray o GMO Detection
b. Medical Molecular Biology o PCR based diagnosis of
genetic and infectious diseases o Applications of DNA
typing o Production of recombinant human therapeutic
proteins o Molecular studies on hearing and vision
impairment o Regenerative Medicine (Stem cell therapy)
o Molecular Virology

c. Environmental Biotechnology
 Plant Molecular Biology

TRANSGENIC SEARCH FOR QUALITY SEED

PLANTS DROUGHT PRODUCTION

Rice Cotton Potato .

Maize

Biosafety Sugarcane Gladiolus

Single Gene High Throughput

Search Approach

Gene Differential Mutant cDNA

Homology Display Identification Library

Promoter/Transcripti Microarra
on Factor Studies
Analysis
Transformation of
Crops

Practical # 1
Recipe of MS- media
Ingredients Concentration (g/L)
MS (murashige and skooge) 4.43
Sucrose 30
Phyta gel 3

Procedure
1- Take 1 litter distal water in the flask by measuring cylinder
2- Take 4.43-gram MS media and add into water in flask
3- Add 30-gram sucrose in the flask
 4- Mix it completely by shaker
5- Measure the pH by pH meter (5.7-5.8). If pH is less than 5.7-5.8 then added few drops
of NaOH and if pH is greater than 5.7-5.8 then added few drops of HCl for maintaining
pH (5.7-5.8).
6- After that phyta gel is added in this solution for solidifying it.
7- After shaking, cover the mouth of flask is covered with paper and aluminum foil
8- Then autoclaved at 121 0c for 15 mints.
Practical #02
MS Broth Preparation
Recipe for 1 liter
Ingredients Concentration (g/L)
MS 4.43
Sucrose 30

Procedure for this media is same as like the MS- media. But phyta gel is not added into this
media.

Practical # 03

LB (Luria and Bertani) Media Preparation

Recipe for 1 litter media

Ingredients Concentration (g/L)
Trypton 10
NaCl 10
Yeast extract 5
Agar 18

Procedure
1. Take 1 litter distil water by measuring cylinder in autoclaved flask
2. Added 10 gram per liter trypton in it
 3. Then added 10gram sodium chloride After that added 5gram yeast extract in it

4. Now shake it well on magnet shaker.

5. pH is maintained at 7.5, by adding HCL (for decreasing) and NaCl (for increasing). pH
meter used, is calibrated by already prepared solution with pH of 4. If meter is giving
the reading of 4 it means it is ready to use.
6. In the last, added 18gram agar for solidifying this material.
7. After shaking, cover the mouth of flask is covered with paper and aluminum foil
8. Then autoclaved at 1210c for 15 mints.
Before cooling it, pour onto plates for future use. Pouring is done in cabinet system to avoid
contamination.

Practical # 04
LB (liquid Broth) Media Preparation
Recipe for 1 liter
Ingredients Concentration (g/L)
Tryptone 10
NaCl 10
Yeast extract 5

For this media, preparation method is same as like the Luria and Bertani but in this media agar
is not used.

Practical # 05
Callus Media Preparation
Recipe for one liter
Ingredients Concentration (g/L)
MS 4.43
Sucrose 30
Myoinsetol 0.25
Casin 1
Phyta gel 3.7

Procedure
1- Take 1 litter distal water in the flask by measuring cylinder
2- Take 4.43-gram MS media and add into water in flask
3- Added 30-gram sucrose in the flask
4- Added myoinsetol and casin (0.25 and 1gram)
5- Then added suitable concentration of 2,4-D in it.
6- mixed it by the help of magnetic shaker
7- then pH is maintained at 7.5, by adding HCL (for decreasing) and NaCl (for
increasing). pH meter used, is calibrated by already prepared solution with pH of 4. If
meter is giving the reading of 4 it means it is ready to use.
8- After that added phyta gel to about 3.7gram
9- After shaking, cover the mouth of flask is covered with paper and aluminum foil
9. Then autoclaved at 1210c for 15 mints.
Before cooling it, pour onto plates for future use. Pouring is done in cabinet system to avoid
contamination.