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Isolation, characterization and expression of a novel vegetative insecticidal protein gene of Bacillus thuringiensis (2005)

Bhalla, Ritu ; Dalal, Monika ; Panguluri, Siva K. ; [et al.]

Oxford, UK : Blackwell Publishing Ltd

FEMS microbiology letters 243 (2005), S. 0

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ISSN: 1574-6968

Source: Blackwell Publishing Journal Backfiles 1879-2005

Topics: Biology

Notes: Twenty-four serovars of Bacillus thuringiensis (Bt) were screened by polymerase chain reaction to detect the presence of vegetative insecticidal protein gene (vip)-like sequences by using vip3Aa1-specific primers. vip-like gene sequences were identified in eight serovars. These genes were cloned and sequenced. The deduced amino acid sequence of the vip3Aa14 gene from Bacillus thuringiensis tolworthi showed considerable differences as compared to those of Vips reported so far. The vip3Aa14 gene from Bt tolwarthi was expressed in Escherichia coli using expression vector pET29a. The expressed Vip3Aa14 protein was found in cytosolic supernatant as well as pellet fraction, but the protein was more abundant in the cytosolic supernatant fraction. Both full-length and truncated (devoid of signal sequence) Vips were highly toxic to the larvae of Spodoptera litura and Plutella xylostella. Truncation of Vip3Aa14 protein at N-terminus did not affect its insecticidal activity.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1016/j.femsle.2005.01.011

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AFLP fingerprinting and genotypic characterization of some serovars of Bacillus thuringiensis (2000)

Pattanayak, Debasis ; Srinivasan, K. ; Mandaokar, Ajin D. ; [et al.]

Springer

World journal of microbiology and biotechnology 16 (2000), S. 667-672

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ISSN: 1573-0972

Keywords: AFLP ; Bacillus thuringiensis ; DNA fingerprinting ; genetic diversity

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: Abstract Amplified fragment length polymorphism (AFLP)-based fingerprinting of 24 serovars of Bacillus thuringiensis (Bt) representing different serotypes was performed using 13 EcoR1 (+2) and Mse1 (+3) primer combinations for genotypic characterization. A high degree of polymorphism was established among the Bt serovars. A total of 1107 fragments ranging from 30–850 bp were generated out of which 1106 were polymorphic. Discrimination rates of different primer combinations at various band levels (1–5) among different Bt serovars were more than 90%. Cluster analysis revealed very low similarity values, ranging from 7–50%, among the Bt serovars indicating their remarkable genetic diversity. AFLP analysis establishes the molecular relatedness between the serovars and serotypes.