Characterization of a Brassica napus gene encoding a cruciferin subunit: estimation of sizes of cruciferin gene families

Rödin, Joakim; Sjödahl, Staffan; Josefsson, Lars‐Göran; Rask, Lars

doi:10.1007/bf00040615

Cited by 23 publications

(16 citation statements)

References 20 publications

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“…RNA was transferred to Hybond C-extra membrane (Amersham) according to the manufacturer's instructions. The Southem filter with genomic DNA was as described by Rodin et al (1993). Both Southern and northern filters were hybridized (Sambrook et al, 1989) in 6X SSC (1X SSC is 0.15 M NaCl, 0.015 M sodium citrate), 5X Denhardt's solution, 0.1% SDS, 100 pg/mL single-stranded salmon sperm, with the PCR fragment described above as a probe (see "Cloning and Sequence Analysis") and washed in 2X SSC, 0.1% SDS at room temperature followed by 1 X SSC, O.lo/0 SDS, at 65°C for 30 to 60 min (northern blot for seed time curve and Southern blot) or 2X SSC, 0.1% SDS at 65°C for 30 min (northern blot for tissue distribution and induction studies).…”

Section: Northern and Southern Hybridizationsmentioning

confidence: 99%

A Wound- and Methyl Jasmonate-Inducible Transcript Coding for a Myrosinase-Associated Protein with Similarities to an Early Nodulin

Taipalensuu¹,

Falk²,

Rask³

1996

Plant Physiol.

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Myrosinase is regarded as a defense-related enzyme in the Brassicaceae and is capable of hydrolyzing glucosinolates into various compounds, some of which are toxic. Severa1 myrosinase isoenzymes exist, and some of them have been found in association with nonmyrosinase proteins. One of these associated proteins, myrosinase-associated protein (MyAP), was purified from seeds of Brassica The myrosinase-glucosinolate system is a preformed, two-component system that is activated upon tissue damage, whereby enzymatic decomposition of glucosinolates by the myrosinase enzyme takes place. Glucosinolates and their hydrolysis products are claimed to function as a defense against generalized herbivores and are implicated as a factor in host-plant recognition by specialized consumers (Chew, 1988;Louda and Mole, 1991). Myrosinase (thioglucoside glucohydrolase, EC 3.2.3.1) consists of a group of isoenzymes capable of hydrolyzing the thioglucoside bond in glucosinolates. The resulting aglycones rearrange spontaneously and yield, in addition to sulfate, compounds such as isothiocyanates, organic thiocyanates, epithionitriles, and nitriles. Within individual plants of a species, several different types of glucosinolates exist. The exact outcome of the reaction depends on a number of factors '

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Section: Northern and Southern Hybridizationsmentioning

confidence: 99%

A Wound- and Methyl Jasmonate-Inducible Transcript Coding for a Myrosinase-Associated Protein with Similarities to an Early Nodulin

Taipalensuu¹,

Falk²,

Rask³

1996

Plant Physiol.

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“…Lines expressing a moderate level of barnase were selected for further studies. Expression of barnase2 A. Two-insert segregating double RBF containing two blocking constructs encoding two identical barnase1 mRNA and driven by different promoters: cysteine endopeptidase (SH-EPp) from Vigna mungo (Akasofu et al, 1990) and cruciferin promoter (CRUp) from Brassica napus (Rodin et al, 1992). The GUS gene models the transgene of interest.…”

Section: One-insert Double Rbfmentioning

confidence: 99%

“…Abbreviations: NTS: Nicotiana tabacum cv. Samsung; HSp: heat shock promoter of soybean (Czarnecka et al, 1989); SH-EPp: cysteine endopeptidase promoter from Vigna mungo (Akasofu et al, 1990); CRUp: cruciferin promoter of Brassica napus (Rodin et al, 1992). Several lines of transgenic tobacco plants carrying the insert with non-segregating RBF (Fig.…”

Section: One-insert Double Rbfmentioning

confidence: 99%

“…The blocking construct used in our previous study (Kuvshinov et al, 2001; consisted of a barnase gene, regulated by the seed germination specific cysteine endopeptidase promoter (Akasofu et al, 1990;Yamauchi et al, 1996), which is specifically active during embryogenesis and seed germination (Kuvshinov et al, 2001). We have also used a cruciferin promoter originating from Brassica napus, which is active during embryogenesis (Rodin et al, 1992). The RC consisted of a barstar gene driven by the soybean heat shock promoter (Czarnecka et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

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Double recoverable block of function – a molecular control of transgene flow with enhanced reliability

Kuvshinov¹,

Anisimov²,

Yahya³

et al. 2005

Environ. Biosafety Res.

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Despite all the achieved benefits and potential promises from recombinant DNA technology of plants, the potential of transgene spread to wild relatives and to non-transgenic crops is still of a wide-spread concern. We continue to develop recoverable block of function (RBF) technology for gene flow control in transgenic plants. RBF consists of two elements: blocking construct (BC) and recovering construct (RC). Natural expression of the BC (barnase) in embryos and sprouts blocks a physiological function essential for survival or reproduction of the transgenic plant (mRNA synthesis and germination). Artificially induced (heat shock treatment) RC (barstar) recovers the blocked function enabling transgenic plant to reproduce. In natural conditions without artificial induction of RC the transgenic plant can not reproduce itself. However, a single RBF may still fail because of the potential for mutations and gene silencing of the inserted constructs. To minimize the frequency of such an inactivation, we developed a double RBF, in which a single insert comprising two BC, flanking a transgene of interest, was constructed and transferred into tobacco (Nicotiana tabacum (L.)). We used a barstar gene driven by a heat shock or 35S promoter as a RC, and two different promoters were used for barnase genes in the BC. One BC contained a seed germination specific cysteine endopeptidase promoter (BC 1 ) and the other contained the cruciferin promoter (BC 2 ), which is active during fruit development and embryogenesis. Three alternative constructs of double RBF are described, and a segregating two-insert as well as a one-insert cassettes, were compared. One-insert system comprising two BC with different nucleotide sequences but degenerate codons that expressed the same Barnase protein appeared to be the most reliable choice. The biological and molecular data obtained suggest that double RBF is a potent transgene containment technique that can safely be applied in agriculture.

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“…Southern blots with 10 to 20 p g of restriction-enzymedigested genomic DNA from the dihaploid line 20516 K were as described by Rodin et al (1993). Isolation of total RNA and northern blotting with the Hybond C Extra membrane (Amersham) were performed as described (Falk et al, 1992).…”

Section: Southern and Northern Hybridizationsmentioning

confidence: 99%

Expression of a Zeatin-O-Glucoside-Degrading [beta]-Glucosidase in Brassica napus

Falk

Rask

1995

Plant Physiol.

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Characterization of a Brassica napus gene encoding a cruciferin subunit: estimation of sizes of cruciferin gene families

Cited by 23 publications

References 20 publications

A Wound- and Methyl Jasmonate-Inducible Transcript Coding for a Myrosinase-Associated Protein with Similarities to an Early Nodulin

A Wound- and Methyl Jasmonate-Inducible Transcript Coding for a Myrosinase-Associated Protein with Similarities to an Early Nodulin

Double recoverable block of function – a molecular control of transgene flow with enhanced reliability

Expression of a Zeatin-O-Glucoside-Degrading [beta]-Glucosidase in Brassica napus

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