Indian Journal of Experimental Biology

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VOLUME 49

NUMBER 7

JULY 2011

CODEN: IJEB (A6) 49 (7) 475–560 (2011)

ISSN: 0019-5189 (print); 0975-1009 (online)

CONTENTS

 

Papers

 

 

 

Effect of anti-fat body antibodies on reproductive capacity of mosquito Anopheles stephensi and transmission blocking of Plasmodium vivax

479

Monika Gulias-Nuss, Aditya Mundhalia & Surendra K Gakhar

 

 

 

Chemomodulatory effect of Dolichos biflorus Linn. on skin and forestomach papillomagenesis in Swiss albino mice

483

Rajesh Nanta & Raosaheb K Kale

 

 

 

Effect of silymarin on curcumin-induced mortality in zebrafish (Danio rerio) embryos and larvae

491

Rong-Jen  Shiau, Pei-Chun Shih & Yu-Der Wen

 

 

 

Protective effect of aqueous garlic extract against lead-induced hepatic injury in rats

498

Debamita Kilikdar, Debasri Mukherjee, Elina Mitra, Arnab K Ghosh, Anjali Basu,
Ananga Mohan Chandra & Debasish Bandyoapdhyay 

 

 

 

Protective effect of kombucha tea against tertiary butyl hydroperoxide induced cytotoxicity and cell death in murine hepatocytes

511

Semantee Bhattacharya, Prasenjit Manna, Ratan Gachhui & Parames C Sil

 

 

 

Antiulcer activity of Andrographis paniculata (Burm.f.) Wall. against cysteamine-induced duodenal ulcer in rats

525

P  Saranya & A Geetha

 

 

 

Effect of Ayurvedic mercury preparation Makarsdhwaja on geriatric canine —
A preliminary study

534

S Sinyorita, C K Ghosh, A Chakrabarti, B Auddy, Runa Ghosh & P K Debnath

 

 

 

Combined effect of dark and wounding on regeneration potential of Houttuynia cordata Thunb. leaves

540

Y Wen Xu, Jian Wei Zengh, Yu Ting Zou, Amjad M Husaini, Ru Yu Yao,
De Gang Wu & Wei Wu

 

 

 

 

..contd

Distinct synergistic action of piperacillin and methylglyoxal against
Pseudomonas aeruginosa

547

Sayanti Mukherjee, Shaswati Chaki, Sukhien Das, Saswati Sen, Sasmir Kr Dutta &
Sujata G Dastidar

 

 

 

Notes

 

 

 

Inhibition of Naja nigricolis (Reinhardt) venom protease activity by Luffa egyptiaca (Mill) and Nicotiana rustica (Linn) extracts

552

M A Ibrahim, A B Aliyu, A Abusufiyanu, M Bashir & A B Sallau

 

 

 

Behavioural responses of desert gerbil, Meriones hurrianae after removal of scent marking gland

555

Mohd. Idris & R S Tripathi

 

 

 

A novel method of plasmid isolation using laundry detergent

558

P Yadav, A Yadav, V Garg, T K Datta, S L Goswami & S De

 

 

 

 

Editor’s Note

The Indian Journal of Experimental Biology is covered by the following international abstracting and indexing services:

 

Science Citation Index ExpandedTM

PubMed (http://www.ncbi.nim.nih.gov/)

MEDLINE

BIOSIS

Chemical Abstracts Service

Excerpta Medica

Informascience

Refrativnyi Zhurnal

Zoological Records

 

_____________________

 

Indian Journal of Experimental Biology in Open Access Mode

 

  The Indian Journal of Experimental Biology (IJEB) is now an open access journal in the repository, NISCAIR Online Periodicals Repository (NOPR) [http://nopr.niscair.res.in].

  Full text of all articles published in IJEB from 2006 onwards can now be accessed at NOPR in the open access mode. Papers in the current issue shall be uploaded immediately. Papers published in earlier years shall be added soon.

  NOPR is based on DSpace, a digital repository software, and allows document browsing, document searching and various search options like title, author name, keywords, year, issue, etc.

 

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Author Index

Abusufiyanu A

552

Aliyu A B

552

Auddy B

534

 

 

Bandyoapdhyay Debasish

498

Bashir M

552

Basu Anjali

498

Bhattacharya Semantee

511

 

 

Chaki Shaswati

547

Chakrabarti A

534

Chandra Ananga Mohan

498

 

 

Das Sukhen

547

Dastidar Sujata G.

547

Datta T K

558

De S

558

Debnath P K

534

Dutta Samir Kr

547

 

 

Gachhui Ratan

511

Gakhar Surendra K

479

Garg V

558

Geetha A

525

Ghosh Arnab K

498

Ghosh C K

534

Ghosh Runa

534

Goswami S L

558

Gulia-Nuss Monika

479

 

 

Husaini Amjad M

540

 

 

Ibrahim M A

552

Idris Mohd.

555

 

 

Kale Raosaheb K

483

Kilikdar Debamita

498

 

 

Manna Prasenjit

511

Mitra Elina

498

Mukherjee Debasri

498

Mukherjee Sayanti

547

Mundhalia Aditya

479

 

 

Nanta Rajesh

483

 

 

Sallau A B

552

Saranya P

525

Sen Saswati

547

Shiau Rong-Jen

491

Shih Pei-Chun

491

Sil Parames C

511

Sinyorita S

534

 

 

Tripathi R S

555

 

 

Wen Yu-Der

491

Wu De Gang

540

Wu Wei

540

 

 

Xu Y Wen

540

 

 

Yadav A

558

Yadav P

558

Yao Ru Yu

540

 

 

Zeng Jian Wei

540

Zou Yu Ting

540

 

 

Keyword Index

Andrographis paniculata

525

Antimicrobial action

547

Anti-mosquito antibodies

479

Antioxidant

498,

 

511

Ayurvedic mercury preparation

534

 

 

Basolateral membrane

525

Benzo(a)pyrene

483

Black tea

511

Brushborder membrane

525

 

 

Cell death

511

Chemomodulation

483

Curcumin

491

Cysteamine

525

Cytioprotection

511

Cyto-toxicity

511

 

 

Dark

540

Desert gerbil

555

Dolichos biflorus

483

Duodenal ulcer

525

 

 

Fat body

479

 

 

Garlic

498

Geriatric dog

534

 

 

Hepataoprotection

491

Hepatoacytes

511

Houttuynia cordata

540

 

 

Kombucha tea

511

 

 

Laundry detergent

558

Lead

498

Luffa egyptiaca

552

 

 

Makaradhwaja

534

Malaria

479

Meriones hurrianae

555

Methylglyoxal

547

Mucin

525

 

 

Naja nigricolis

552

Nicotiana rustica

552

 

 

Oxidative stress

498

 

 

Papillomagenesis

483

Peroxidative damage

483

Piperacillin

547

Plasmid

558

Plasmodium

479

Protease

552

Pseudomonas aeruginosa

547

 

 

Reactive oxygen species

511

Regeneration

540

 

 

Scent marking behaviour

555

Synergism

547

 

 

Tertiary butyl hydroperoxide

511

Transmission blocking

479

 

 

Vector

558

Venom

552

 

 

Wounding

540

 

 

Zebrafish

491

 

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 479-482

 

 

Effect of anti-fat body antibodies on reproductive capacity of mosquito Anopheles stephensi and transmission blocking of Plasmodium vivax

Monika Gulia-Nuss 1*, Aditya Mundhalia 2 & Surendra K Gakhar 3

1 Department of Entomology, University of Georgia , Athens , GA 30602 , USA

2 Department of Biotechnology, Career Institute of Technology and Management, Faridabad 121 001 India

3 Advance Centre for Biotechnology, Maharshi Dayanand University , Rohtak 12 4001, India

 

Received 14 May 2010; revised 6 March 2011

Effect of anti-mosquito-fat body antibodies on the development of the malaria parasite, Plasmodium vivax has been studied by feeding Anopheles stephensi mosquitoes with infected blood supplemented with serum from immunized rabbits. Immunogenic polypeptides were identified by western blot. Mosquitoes that ingested anti-fat body antibodies along with infectious blood meal had significantly fewer oocysts than the mosquitoes in the control group. Effect of anti-mosquito fat body antibodies on fecundity, hatchability, mortality and engorgement of mosquitoes has also been reported. A significant reduction in fecundity and hatchability was observed, however, effect on mortality and engorgement was variable and statistically insignificant. Results indicated that fat body antibodies have the potential to disrupt reproductive physiology of malaria vector An. stephensi .

 

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 483-490

 

 

Chemomodulatory effect of Dolichos biflorus Linn. on skin and forestomach papillomagenesis in Swiss albino mice

Rajesh Nanta* & Raosaheb K Kale

School of Life Sciences , Jawaharlal Nehru University , 110 067, New Delhi , India

Received 6 September 2010; revised 3 March 2011

Effect of consumption of three different doses (2%, 4% and 6%, w/w) of Dolichos biflorus Linn. seeds on hepatic drug metabolizing enzymes, antioxidant enzymes, reduced glutathione content, lactate dehydrogenase and lipid peroxidation in Swiss albino mice has been reported. Anti-carcinogenic effect has been studied by 7,12-dimethylbenzanthracene (DMBA)-induced skin and benzo(a)pyrene[B(a)P]-induced forestomach papillomagenesis models. D. biflorus consumption resulted in a significant increase in hepatic carcinogen metabolizing enzyme systems especially at 4% and 6% doses. Significant increase in reduced glutathione content (GSH) and specific activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GR) in liver of mice, at 4% and 6% doses has been reported. Lactate dehydrogensae (LDH) activity and peroxidative damage has been significantly decreased at 4% and 6% doses. In skin papillomagenesis model, 4% and 6% dose in diet significantly reduced the tumor incidence
(up to 25%), tumor multiplicity (up to 59%) and tumor volume per mouse (up to 70%) as compared to DMBA treated group. Importantly, significant reduction in tumor incidence (up to 33%) and tumor multiplicity (up to 61%) was evident for forestomach papillomagenesis model.

 

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 491-497

 

 

Effect of silymarin on curcumin-induced mortality in zebrafish ( Danio rerio ) embryos and larvae

Rong-Jen Shiau 1 , Pei-Chun Shih 2 & Yu-Der Wen 2 *

1 Department of Beauty Science, Chien kuo Technology University , Changhua 50058 , Taiwan

2 Department of Biology, National Changhua University of Education , Changhua 50058 , Taiwan

Received 4 November 2010; revised 20 April 2011

In presence of 7.5 m M of curcumin, no embryos or larva of zebrafish survived 3 days of incubation; however, co-incubation with 144 m g/ml silymarin increased the survival rates of curcumin-treated embryos and larvae to about 70%. Moreover, in presence of 12.5 m M curcumin, all embryos died after 2 days of incubation; however, co-treatment with 144 m g/ml silymarin increased the survival rates of curcumin-treated embryos and larvae up to 60 and 50%, respectively. This protective effect was not found in the other phenolic compounds viz., ferulic acid, naringin, or crocin, tested. Finally, using a fluorescence microscope, accumulation of less curcumin has observed in the edema sac area of the larvae co-treated with curcumin and silymarin than in the larvae treated with curcumin only. The result suggests that the protective effects of silymarin may be due to a decreased accumulation of curcumin in the fish body.

 

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 498-510

 

 

Protective effect of aqueous garlic extract against lead-induced
hepatic injury in rats

Debamita Kilikdar, Debasri Mukherjee, Elina Mitra, Arnab K Ghosh, Anjali Basu, Ananga
Mohan Chandra & Debasish Bandyoapdhyay*

Oxidative Stress and Free Radical Biology Laboratory, Department of Physiology,

University of Calcutta , 92, APC Road , Kolkata 700 009, India

Received 14 September 2010, revised 11 March 2011

Effect of aqueous extract of garlic on hepatic injury due to lead-induced oxidative stress in experimental rats has been investigated. Lead acetate (LA) at a dose of 15 mg / kg body wt was administered ip to rats for 7 consecutive days to induce hepatic injury. Freshly prepared aqueous garlic extract (AGE) at a dose of 50 mg /kg body wt was fed orally to rats 1 h before LA treatment for similar period. LA treatment caused hepatic injury as evident from increased activities of serum glutamate pyruvate transaminase (SGPT) and alkaline phosphatase (ALP), increased serum bilirubin level and damage in the tissue morphology. Lead-induced oxidative stress in liver was evident from increased levels of lipid peroxidation and reduced glutathione. The decreased activity of superoxide dismutase (SOD) and an increased activity of catalase as well as an increased activity of xanthine oxidase (XO) indicate generation and possible accumulation of reactive oxygen intermediates. Furthermore, altered activities of lactate dehydrogenase (LDH), isocitrate dehydrogenase (ICDH), alpha-keto glutarate dehydrogenase (a-KGDH) and succinate dehydrogenase (SDH) also indicate an impaired substrate utilization and generation of oxidative stress. All these changes were found to be mitigated when the rats were pre-treated with the AGE. Results indicate that AGE has the potential to ameliorate lead-induced hepatic injury due to oxidative stress in rats.
The protective effects may be due to the antioxidant properties of AGE and may have future therapeutic relevance.

 

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 511-524

 

 

 

Protective effect of kombucha tea against tertiary butyl hydroperoxide induced cytotoxicity and cell death in murine hepatocytes

Semantee Bhattacharya a , Prasenjit Manna b , Ratan Gachhui a & Parames C Sil b*

a Department of Life Sciences and Biotechnology, Jadavpur University , 188 Raja S C Mullick Road, Kolkata 700 032, India

b Division of Molecular Medicine, Bose Institute, P-1/12 CIT Scheme VII M, Kolkata 700 054, India

Received 17 August 2010; revised 29 March 2011

Kombucha (KT), a fermented black tea (BT), is known to have many beneficial properties. In the present study, antioxidant property of KT has been investigated against tertiary butyl hydroperoxide (TBHP) induced cytotoxicity using murine hepatocytes. TBHP, a reactive oxygen species inducer, causes oxidative stress resulting in organ pathophysiology. Exposure to TBHP caused a reduction in cell viability, increased membrane leakage and disturbed the intra-cellular antioxidant machineries in hepatocytes. TBHP exposure disrupted mitochondrial membrane potential and induced apoptosis as evidenced by flow cytometric analyses. KT treatment, however, counteracted the changes in mitochondrial membrane potential and prevented apoptotic cell death of the hepatocytes. BT treatment also reverted TBHP induced hepatotoxicity, however KT was found to be more efficient. This may be due to the formation of antioxidant molecules like D-saccharic acid-1,4-lactone (DSL) during fermentation process and are absent in BT. Moreover, the radical scavenging activities of KT were found to be higher than BT. Results of the study showed that KT has the potential to ameliorate TBHP induced oxidative insult and cell death in murine hepatocytes more effectively than BT.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 525-533

 

 

Antiulcer activity of Andrographis paniculata (Burm.f.) Wall. against cysteamine-induced duodenal ulcer in rats

P Saranya & A Geetha*

Department of Biochemistry, Bharathi Women's College (Affiliated to University of Madras ), Chennai 600 108, India

Received 10 January 2011; revised 9 March 2011

Antiulcer activity of Andrographis paniculata was evaluated by cysteamine induced duodenal ulcer model in rats. Male albino Wistar rats were pre-administered with 200 mg/kg body wt. of hydroalcoholic extact of Andrographis paniculata (HAEAP) orally, for 30 days prior to i.p. administration of 420 mg/kg body wt. of cysteamine as a single dose. Rats pre-administered with 30 mg/kg body wt. of ranitidine served as standard drug. Ulcer index, thiobarbituric acid reactive substances, mucin, glutathione peroxidase and myeloperoxidase activities, reduced glutathione/oxidized glutathione (GSH/GSSG) ratio, glycoproteins and membrane bound enzyme activities were measured in duodenum of experimental animals. The ulcer score and myeloperoxidase activity were significantly minimized in rats treated with HAEAP. Mucin content was found to be preserved in rats treated with the extract. GSH/GSSG ratio and glutathione peroxidase activities were found to be maintained by the HAEAP. Level of lipid peroxidation products was found to be significantly low in HAEAP treated rats compared to ulcer control rats. The basolateral and brush border membrane bound enzyme activities which were depleted significantly in ulcer control rats were found to be maintained in rats pre-treated with the extract. The ulcer preventing effect was comparable to that of ranitidine treated rats. Level of glycoproteins was also found to be preserved in rats treated with the extract. The normal rats treated with the HAEAP did not show any abnormal alterations in the parameters studied. Histopathological observations also showed the ulcer preventing effect of the HAEAP. It is suggested that the ulcer preventing effect may be due to its mucin preserving and antioxidant nature.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 534-539

 

 

Effect of Ayurvedic mercury preparation Makaradhwaja on
geriatric canine–A preliminary study

S Sinyorita 1 , C K Ghosh 1 , A Chakrabarti 1 , B Auddy 2 , Runa Ghosh 2 & P K Debnath 3*

1 Department of Veterinary Medicine, Ethics and Jurisprudence, West Bengal University of Animal and Fishery Sciences,
Kolkata 700 037, India 2 Natreon Inc. Ltd, Kolkata 700 091, India 3 National Research Institute of Ayurveda for
Drug Development, Kolkata 700 091, India

Received 22 June 2010; revised 30 March 2011

Makaradhwaja , an alchemical Ayurvedic mercury preparation is used as stimulant and vitalizer. Towards veterinary practices, the acceptability, tolerability and toxicity studies were undertaken in geriatric pet dogs aged more than 10 years irrespective of breed and sex for future use. Makaradhwaja (2.5 mg/kg) was used with honey once daily for 30 days. Before and after treatment, blood was collected for hematological studies as well as liver, kidney function and anti-oxidant activity. In control group, honey itself showed no appreciable change whereas, Makaradhwaja lowered neutrophil and total leucocyte count. Serum cholesterol, urea, glucose, alanine amino transferase, aspartate amino transferase, sodium, phosphorus and calcium were decreased. Haemoglobin and serum creatinine were significantly increased. There was appreciable physical, behavioral and body weight change including quality of life. The dose was used in replication of human dose (125 mg/50kg). Anti-oxidant study showed significant increase of lipid per oxidation in experimental group while the values of ABTS radical cation decolorisation assay although decreased but did not show any significant changes. Decrease of serum urea and increase of serum creatinine could not be explained on single dose response. Different dose study could only explain the optimum dose to be required in canine practices.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 540-546

 

 

Combined effect of dark and wounding on regeneration potential of Houttuynia cordata Thunb. leaves

Y. Wen Xu 1 , Jian Wei Zeng 1 , Yu Ting Zou 1 , Amjad M Husaini 2 , Ru Yu Yao 1 , De Gang Wu 1 & Wei Wu 1 *

1 Agronomy College , Sichuan Agricultural University, Ya'an 625014, PR China

2 Division of Plant Breeding and Genetics, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir,
Srinagar, Jammu & Kashmir 191 121, India

Received 8 November 2010; revised 7 April 2011

Houttuynia cordata is one of the most potential medicinal and edible wild herb whose resources have decreased sharply due to excessive exploitation. Besides its slow agamic propagation, problems of browning and non-dedifferentiation have prevented the application of micropropagation in H. cordata . Through 4 week pre-culture in darkness and wounding after 1 week pre-culture, the browning rate of leaf explants decreased significantly and resulted in efficient regeneration (20.64 ± 5.94 adventitious buds per explant) on the induction medium. The protocol shall facilitate conservation and commercial cultivation of the endangered species.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 547-551

 

 

Distinct synergistic action of piperacillin and methylglyoxal against
Pseudomonas aeruginosa

Sayanti Mukherjee a , Shaswati Chaki a , Sukhen Das b , Saswati Sen c , Samir Kr. Dutta c & Sujata G. Dastidar a*

a Department of Microbiology, Herbicure Healthcare Bio-Herbal Research Foundation, 7&8 Metro Garden City,
D.H. Road, Pailan, Kolkata 700 104, India

b Department of Physics, Jadavpur University , Kolkata 700 032, India

c Division of Drug Development/Diagnostics and Biotechnology, Indian Institute of Chemical Biology, (IICB) CSIR,
Kolkata 700 032, India

Received 12 October 2010, revised 28 March 2011

The dicarbonyl compound methylglyoxal is a natural constituent of Manuka honey produced from Manuka flowers in New Zealand . It is known to possess both anticancer and antibacterial activity. Such observations prompted to investigate the ability of methylglyoxal as a potent drug against multidrug resistant Pseudomonas aeruginosa . A total of 12 test
P. aeruginosa strains isolated from various hospitals were tested for their resistances against many antibiotics, most of which are applied in the treatment of P. aeruginosa infections. Results revealed that the strains were resistant to many drugs at high levels, only piperacillin, carbenicillin, amikacin and ciprofloxacin showed resistances at comparatively lower levels. Following multiple experimentations it was observed that methylglyoxal was also antimicrobic against all the strains at comparable levels. Distinct and statistically significant synergism was observed between methylglyoxal and piperacillin by disc diffusion tests when compared with their individual effects. The fractional inhibitory concentration index of this combination evaluated by checkerboard analysis, was 0.5, which confirmed synergism between the pair. Synergism was also noted when methylglyoxal was combined with carbenicillin and amikacin.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 552-554

 

 

Inhibition of Naja nigricolis (Reinhardt) venom protease activity by Luffa egyptiaca (Mill) and Nicotiana rustica (Linn) extracts

M A Ibrahim 1 *, A B Aliyu 2 , A Abusufiyanu 1 , M Bashir 1
& A B Sallau 1

Departments of Biochemistry 1 and Chemistry 2 ,
Ahmadu Bello University , Zaria , Nigeria

Received 9 August 2010; revised 26 April 2011

Luffa egyptiaca and Nicotiana rustica are used in traditional medicine to treat snakebites and were evaluated for inhibitory activities on Naja nigricolis venom protease. The aqueous and ethanolic extracts of L. egyptiaca significantly reduced the maximum velocity (V max ) and the computed index of physiological efficiency ( K cat ) of the enzyme in a dose dependent fashion. The protease activity was non-competitively inhibited by the aqueous extract of N. rustica with the V max significantly decreased and the K M remained unchanged. However, the N. rustica ethanol extract completely inhibited the protease activity. Ethyl acetate fractions partitioned from ethanol extracts of both plants were also found to completely inhibit the N. nigricolis venom protease activity at 0.1 and 0.05%. The use of these plants could be important in the treatment of snakebites.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 555-557

 

 

Behavioural responses of desert gerbil, Meriones hurrianae after removal of
scent marking gland

Mohd. Idris* & R S Tripathi

All India Network Project on Rodent control, Central Arid Zone Research Institute, Jodhpur 342 003, India

Received 15 December 2010; revised 28 March 2011

The desert gerbil, M. hurrianae scent marks the general substratum in its territory with the sebum exudation of mid abdominal gland and urine. Having assessed number of functions, which scent marking plays in the social life of these rodents, the scent marking behaviour was studied in animals, in which the gland was surgically removed and was compared with that of intact rodents. After recovery from the operation, the scent marking frequency of both male and female M.hurrianae declined significantly and was maintained at a low level. Surprisingly, scent marking with urine also declined considerable with time. After 5 months of the operation, desert gerbils were given a choice to respond to male and female sebum odours. The frequency of their scent marking with either sebum or urine did not show any significant enhancement as compared to their initial marking rate. However, the duration of their stay and scent marking frequency near the source of the sebum odour was more that in the clean side of the cage. The role of such altering behaviors of M. hurrianae and their impact on social organization are discussed.

 

 

Indian Journal of Experimental Biology

Vol. 49, July 2011, pp. 558-560

 

 

A novel method of plasmid isolation using laundry detergent

P Yadav 1 , A Yadav 2 , V Garg 3 , T K Datta 1 , S L Goswami 1 , & S De 1* ,

1 Animal Biotechnology Centre and 2 Animal Biochemistry
Division, National Dairy Research Institute, Karnal 132 001, India .
3 Department of Biosciences and Biotechnology, Banasthali
University , Banasthali 304 022, India .

Received 1 April 2009; revised 1 February 2011

Since the discovery of plasmid, various methods have been developed to isolate plasmid DNA. All the methods have one common and important target of isolating plasmid DNA of high quality and quantity in less time. These methods are not completely safe because of use of toxic chemicals compounds. The developed protocol for plasmid extraction is based on the alkaline lysis method of plasmid preparation (extraction at p H 8.0) with slight modifications. Cell lysis reagent sodium dodecyl sulfate is replaced by lipase enzyme present in laundry detergent. A good plasmid preparation can be made, which is well suited for subsequent molecular biology applications. By taking safety measures on count, contaminants like, RNA and protein can be completely avoided with maximized plasmid yield. The resultant plasmid quality and quantity can be well comparable to other prevalent methods.