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Isolation and preliminary sequence characterization of beta-amylase gene promoters in rye [Secale cereale L.]

100%

Sadowski J., Rorat T., Cooke R., Delseny M.

Journal of Applied Genetics

1997

tom 38

nr 3

The isolation of rye ß-Amy1 and ß-Amy2 gene promoters from nuclear DNA using the inverse polymerase chain reaction (IPCR) technique and characterization of their sequences are presented. The conservation of ß-amylasc coding sequences allowed for simultaneous IPCR amplification of two different promoters with primers designed on the basis of the single known cDNA sequence. Two ß-amylasc gene promoters display a low sequence similarity (47%). Beside consensus TATA and CCAAT boxes, other sequence motives common to both promoters were found. In addition, the homology of amino acid sequences of plant ß-amylases available in the database is discussed.

Isolation, cloning and expression of DNA fragments reflecting open reading frames of 1.8 and 4.6 kb RNAs of the I-18 C gene of Chironomus tentans

100%

Borowicz B. P.

Journal of Applied Genetics

2005

tom 46

nr 1

This study, based on the genetic engineering approach, concerns the regulation of the insect gene expression, but it has also a strong significance for plant protection, which is associated with the future possibility to control insect development more effectively, especially in those species that are pests of cultivated plants. The I-18 C gene of Chironomus tentans is activated by heat-shock and the steroid hormone - ecdysone. This gene produces different transcripts (at least four) by alternative splicing. Two different open reading frames, i.e. ORFs I and II of the two main transcripts of this gene, i.e. the 1.8 and 4.6 kb RNA, had been isolated, at the DNA level, as the ORFs reflecting DNA fragments, using the polymerase chain reaction, then were cloned into the bluescript vector and finally were cloned into the pET-3a vector to be translated in the T7 RNA polymerase/promoter system of E. coli. As the preliminary outcome of these experiments it was possible to obtain ORF I overexpressed as the polypeptide, whereas ORF II was expressed as the polypeptide that was strongly visible on SDS-polyacrylamide gel. This could facilitate further studies regarding the mechanisms of expression of this gene, including the level of expression of this gene during the C. tentans development.

Ograniczanie wyników

2 Journal of Applied Genetics

1 Borowicz B. P.

1 Cooke R.

1 Delseny M.

1 Rorat T.

1 Sadowski J.

1 2005

1 1997

Wyniki wyszukiwania

Isolation and preliminary sequence characterization of beta-amylase gene promoters in rye [Secale cereale L.]

Isolation, cloning and expression of DNA fragments reflecting open reading frames of 1.8 and 4.6 kb RNAs of the I-18 C gene of Chironomus tentans