Abstract Differentiation of Bacillus thuringiensis subspecies using repetitive extragenic palindromic PCR (REP PCR) genomic fingerprinting

As. Pac. J. Mol. Biol. & Biotech., Dec 1999 Vol. 7(1) : 79-84

Differentiation of Bacillus thuringiensis subspecies using repetitive extragenic palindromic PCR (REP PCR) genomic fingerprinting

Krisana Phucharoen1, Wipa Chungjatupornchai2 and Sakol Panyim1,2*

1Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus,
Nakornpathom, Thailand; 1National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok, Thailand.

(Received 29 December 1998 / Accepted 26 April 1999)

Abstract.
A total of 29 subspecies of B. thuringiensis strains previously used to raise 28 standard antisera, were differentiated using PCR fingerprinting with primers derived from the Repetitive Extragenic Palindrome (REP) sequence. Using REP specific primers (REP2-1 and REP3) in a defined PCR condition, a reproducible fingerprinting pattern in the size range of 300 2,000 bp was obtained. The bacterial strains can be distinguished by comparing fingerprint patterns. The dendrogram of the fingerprints revealed that the B. thuringiensis subspecies fell into four major clusters with a level of similarity of approximately 20%. The relatedness of the REP-PCR fingerprint patterns of most B. thuringiensis subspecies did not correlate with the bacterial classification based on flagella serotyping. The REP-PCR fingerprint method should prove to be a rapid and simple tool for numerical taxonomic analysis of B. thuringiensis strains.

Keywords: Bacillus thuringiensis, REP PCR, genomic Fingerprintin

Abstract Production of high frequency multiple shoots and plantlets in six Indian cultivars of cotton (Gossypium hirsutum L.)

As. Pac. J. Mol. Biol. & Biotech., Dec 1999 Vol. 7(1) : 73-78

Production of high frequency multiple shoots and plantlets in six Indian cultivars of cotton (Gossypium hirsutum L.)

A Mhatre, P. Suprasanna, R. Jaiswal, V. V. Kumar and P. S. Rao*

Nuclear Agriculture & Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
Email. This email address is being protected from spambots. You need JavaScript enabled to view it.

(Received 10 December 1998 / Accepted 4 March 1999)

Abstract.
A protocol for the production of high frequency multiple shoots using seedling apices with cotyledonary node of six Indian cultivars of cotton (Gossypium hirsutum L.) has been developed. An average of 7 10 shoots per explant were produced directly from the shoot apices excised from 10 day old seedlings on MS medium containing BAP (8.87?M) along with adenine sulphate (81.43 ?M) or kinetin (4.67 ?M). Rooting of isolated shoots was achieved on MS medium containing NAA (0.054?M). Rooted plantlets exhibited 95% survival during hardening in the greenhouse and regenerated plants grew to maturity with normal flowering and boll set.

Keywords: Cotton, Gossypium hirsutum, plant regeneration, in vitro culture

Abstract Synergistic effects of medium strategies and sucrose levels with nitrogenous compounds, phosphate and myo inositol on Morinda elliptica cell cultures

As. Pac. J. Mol. Biol. & Biotech., Dec 1999 Vol. 7(1) : 61-72

Synergistic effects of medium strategies and sucrose levels with nitrogenous compounds, phosphate and myo inositol on Morinda elliptica cell cultures

M.A. Abdullah1, M. Marziah2, M.N. Zakaria1, A.M. Ali1, N.H. Lajis3 and A.B. Ariff1*

1Department of Biotechnology, 2Department of Biochemistry and Microbiology, 3Department of Chemistry,
Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

(Received 15 October 1998 / Accepted 10 May 1999)

Abstract.
Synergistic effects of medium strategies and sucrose levels with nitrogenous compounds, phosphate and myo inositol on Morinda elliptica cell suspension cultures were studied. Three types of medium strategies were tested maintenance (M), growth (G) and production (P) media. In M medium, incorporation of NO3 between 30 mM and 67.7 mM, was vital for optimum growth and anthraquinone (AQ) production. NO3 level upto 95.9 mM did not affect cell growth though AQ may be slightly reduced. At NO3 level of 77.1 mM, 8% (w/v) sucrose in G medium produced optimum cell growth and AQ content but cell growth was reduced whilst AQ content unaffected in P medium. At 10% (w/v) sucrose in both G and P media, both cell growth and AQ content were reduced. Phosphate was more a growth limiting nutrient than an AQ limiting nutrient. Absence of phosphate retarded cell growth in M medium but no significant difference on cell growth and AQ yield was observed with 5 fold increase in phosphate in M medium; at 8% (w/v) and 10% (w/v) sucrose in G medium; and at 8% (w/v) sucrose in P medium. At 10%(w/v) sucrose in P medium, the AQ yield was reduced by almost 50% (w/v). Nitrogen and phosphate toxicity was not only a function of both individual and collective level of ammonium, nitrate and phosphate, but also the level of sucrose and medium strategies. myo inositol was not an absolute requirement in M. elliptica cell suspension culture.

Keywords: Anthraquiriones, Morinda elliptica, nitrogen, phosphate, myo inosito

Abstract Isolation and characterisation of a cDNA encoding I aminocyclopropane l carboxylate oxidase from mango (Mangifera indica L.)

As. Pac. J. Mol. Biol. & Biotech., Dec 1999 Vol. 7(1) : 53-60

Isolation and characterisation of a cDNA encoding I aminocyclopropane l carboxylate oxidase from mango (Mangifera indica L.)

Z. Zainal*, G. A. Tucker and G. W. Lycett

Department ofPhysiology and Environmental Science, Faculty of Agricultural and Food Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire. LE12 5RD, UK

(Received 15 February 1999 / Accepted 28 May 1999)

Abstract.
A full length cDNA clone designated as pNY601 encoding I aminocyclopropane I carboxylate (ACC) oxidase was isolated from a ripening mango (Mangifera indica L.) cDNA library using pTOM13 as a probe. Plasmid pNY601 is 1213 bp and contains a single open reading frame encoding a 324 amino acid polypeptide with a calculated molecular weight of 36.7 kDa and pI of 5.4. The deduced amino acid sequence shows considerable sequence (80-90%) similarity to other ACC oxidases; from various plant species. Northern blot analysis of RNA isolated from different stages of ripening shows that expression began while the fruits were still at the mature green stage. The appearance of these transcripts before ripening process take place may be due to chilling injury.

Keywords: Gene expression, Mangifiera indica, ACC oxidase, cDNA, fruit ripening

Abstract Use of polymerase chain reaction for improved diagnosis of tuberculosis in children

As. Pac. J. Mol. Biol. & Biotech., Dec 1999 Vol. 7(1) : 29-38

Use of polymerase chain reaction for improved diagnosis of tuberculosis in children

T.H. Tang1, M. Musa1, Z.F. Zainuddin1, K.E. Choo2,L. Mohd Noh1, K A. Myat1,
1. Yaacob1, H. Ismail2, M. Mustaffa3 and Maheran Musa2

1School of Medical Sciences, Universiti Sains Malaysia Cawangan Kelantan, Kota Bharu, Kelantan, Malaysia;
2Paediatric Unit, Kota Bharu Hospital, Kota Bharu, Kelantan, Malaysia; 3Chest Clinic,
Kota Bharu Hospital, Kota Bharu, Kelantan, Malaysia

(Received 8 December 1998 / Accepted 26 February 1999)

Abstract.
Tuberculosis (TB) in children is severe especially for those under 5 years old, with a tendency to extrapulmonary spread to other organs with high morbidity and mortality. Thus, in contrast to most adult cases, TB in children often requires urgent treatment but diagnosis is often difficult to establish by clinical and conventional laboratory methods. Rapid tests based on molecular techniques for diagnosis of TB have been widely reported but relatively few involve paediatric cases. In this study, a polymerase chain reaction (PCR) based test using primers derived from the insertion sequence IS 6110 and a new Mycobacterium tuberculosis complex specific target designated as B9 was developed and used for detecting the organism in specimens taken from four paediatric patients. This method was found to be a useful addition to the current battery of tests available to the paediatrician particularly in cases where diagnoses are difficult to establish.

Keywords: B9, IS6110, Mycobacterium tuberculosis, paediatric, PCR

Sponsors Members

  • image
  • image
  • image
  • 1

About MSMBB

We are a non-profit organisation that was established in 1988 to promote molecular biology and biotechnology.

Stay Connected on:

Contact Us

For general information about MSMBB, including registration, please contact us at:

  Department of Parasitology,
Faculty of Medicine,
University of Malaya,
50603 Kuala Lumpur,
Malaysia.
  This email address is being protected from spambots. You need JavaScript enabled to view it.
  +603 - 7967 4744
  +603 - 7967 4749