Nucleotide sequence of the gene for the<i>Vigna mungo</i>sulfhydryl-endopeptidase (SH-EP)

Akasofu, Harue; Yamauchi, Daisuke; Minamikawa, Takao

doi:10.1093/nar/18.7.1892

Cited by 23 publications

(14 citation statements)

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“…Comparison of the deduced amino acid sequence with protein-sequence databases indicated homology with several plant thiol-protease genes (Fig. 2), especially with VSCYSPROA from common vetch (Becker et al, 1997), vicilin peptide hydrolase from mung bean (Lee et al, 1997), SH-EP from Vigna mungo (Akasofu et al, 1990), EP-C1 from bean (Ogushi et al, 1992), and SEN11 (Guerrero et al, 1998) and SEN102 (Valpuesta et al, 1995) from daylily. The putative thiol-protease activity of the TPE4A protein is supported by the conservation of the three catalytic amino acids of papain, Cys-154, His-298, and Asn-310 (Kamphuis et al, 1985), at positions 153, 288, and 309, respectively (Fig.…”

Section: Molecular Characterization Of Tpe4amentioning

confidence: 96%

“…The alignment was generated using the PileUp program of the Genetics Computer Group package. The compared proteases are: EP-C1 from bean (Ogushi et al, 1992); SEN102 (Valpuesta et al, 1995) from daylily; SEN11 (Guerrero et al, 1998) from daylily; SH-EP from Vigna mungo (Akasofu et al, 1990); vicilin peptide hydrolase from mung bean (VICILIN-PE; Lee et al, 1997); and VS-CYSPROA from vetch (Becker et al, 1998). presenescent ovaries, suggesting a repression of the basal expression levels after the hormonal treatment.…”

Section: Expression Of the Tpe4a Genementioning

confidence: 99%

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Cloning and Characterization ofTPE4A, a Thiol-Protease Gene Induced during Ovary Senescence and Seed Germination in Pea1

1999

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A cDNA clone encoding a thiol-protease (TPE4A) was isolated from senescent ovaries of pea (Pisum sativum) by reverse transcriptase-polymerase chain reaction. The deduced amino acid sequence of TPE4A has the conserved catalytic amino acids of papain. It is very similar to VSCYSPROA, a thiol-protease induced during seed germination in common vetch. TPE4A mRNA levels increase during the senescence of unpollinated pea ovaries and are totally suppressed by treatment with gibberellic acid. In situ hybridization indicated that TPE4A mRNA distribution in senescent pea ovaries is different from that of previously reported thiol-proteases induced during senescence, suggesting the involvement of different proteases in the mobilization of proteins from senescent pea ovaries. TPE4A is also induced during the germination of pea seeds, indicating that a single protease gene can be induced during two different physiological processes, senescence and germination, both of which require protein mobilization.

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Section: Molecular Characterization Of Tpe4amentioning

confidence: 96%

Section: Expression Of the Tpe4a Genementioning

confidence: 99%

Cloning and Characterization ofTPE4A, a Thiol-Protease Gene Induced during Ovary Senescence and Seed Germination in Pea1

1999

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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%

“…To recover the blocked function of the host plant, the expression of the RC is deliberately induced by an external stimulus, such as heat shock. 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).…”

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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Development of gene expression system in egg cells and zygotes isolated from rice and maize

et al. 2017

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Nucleotide sequence of the gene for theVigna mungosulfhydryl-endopeptidase (SH-EP)

Abstract: A genomic clone for SH-EP was isolated from a Vigna mungo EP. Initation and termination codons and the putative

Cited by 23 publications

References 1 publication

Cloning and Characterization ofTPE4A, a Thiol-Protease Gene Induced during Ovary Senescence and Seed Germination in Pea1

Cloning and Characterization ofTPE4A, a Thiol-Protease Gene Induced during Ovary Senescence and Seed Germination in Pea1

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

Development of gene expression system in egg cells and zygotes isolated from rice and maize

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