Reporter gene introduction and transient expression in protoplasts of Porphyra yezoensis

Mizukami, Yuzuru; Kito, Hitoshi; Kunimoto, Masahiko; Murase, Noboru

doi:10.1023/b:japh.0000019050.54703.14

Cited by 18 publications

(11 citation statements)

References 21 publications

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“…For example, hepatitis B surface antigen gene (HbsAg) was expressed in Laminaria japonica (Jiang et al, 2002). However, techniques concerning efficient foreign gene expression in P. yezoensis are still limited (Mizukami et al, 2004). Transient expressions of GUS gene were reported in P. miniata (Kubler et al, 1994) and P. yezoensis (Kuang et al, 1998) under the control of viral CaMV 35 S promoter.…”

Section: Introductionmentioning

confidence: 99%

“…Although 35 S promoter has a wide host range and is widely used in many plant cells (Odell et al, 1985;Jefferson et al, 1987), this virus-derived constitutive promoter is not always the best choice for gene expression due to its tissue specificity or possible toxicity of accumulated target gene product to host cells. Therefore, in order to establish a reliable gene expression system in P. yezoensis, it is important to investigate the usefulness of endogenous promoter sequences (Mizukami et al, 2004). Recently, the promoter sequence of a chloroplast ribuose-bisphosphatecarboxylase /oxygenase (Rubisco) gene has been used for the transient expression in P. yezoensis protoplasts (Mizukami et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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Efficient gusA transient expression in Porphyra yezoensis protoplasts mediated by endogenous beta-tubulin flanking sequences

Gong

Dai

et al. 2007

J Ocean Univ. China

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Endogenous tubulin promoter has been widely used for expressing foreign genes in green algae, but the efficiency and feasibility of endogenous tubulin promoter in the economically important Porphyra yezoensis (Rhodophyta) are unknown. In this study, the flanking sequences of beta-tubulin gene from P. yezoensis were amplified and two transient expression vectors were constructed to determine their transcription promoting feasibility for foreign gene gusA. The testing vector pATubGUS was constructed by inserting 5'-and 3'-flanking regions (Tub5' and Tub3') up-and down-stream of β-glucuronidase (GUS) gene (gusA), respectively, into pA, a derivative of pCAT ® 3-enhancer vector. The control construct, pAGUSTub3, contains only gusA and Tub3'. These constructs were electroporated into P. yezoensis protoplasts and the GUS activities were quantitatively analyzed by spectrometry. The results demonstrated that gusA gene was efficiently expressed in P. yezoensis protoplasts under the regulation of 5'-flanking sequence of the beta-tubulin gene. More interestingly, the pATubGUS produced stronger GUS activity in P. yezoensis protoplasts when compared to the result from pBI221, in which the gusA gene was directed by a constitutive CaMV 35 S promoter. The data suggest that the integration of P. yezoensis protoplast and its endogenous beta-tubulin flanking sequences is a potential novel system for foreign gene expression.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Efficient gusA transient expression in Porphyra yezoensis protoplasts mediated by endogenous beta-tubulin flanking sequences

Gong

Dai

et al. 2007

J Ocean Univ. China

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“…Initially, research was carried out on transient expression of reporter genes under the control of promoters that had been used for similar purposes in higher plants and unicellular algae (Qin et al, 2004). Transient gene expression has been reported in Kappaphycus alvarezii explants (Kurtzman and Cheney, 1991;Wang et al, 2010), Porphyra miniata protoplasts (Kübler et al, 1994), Porphyra yezoensis protoplasts and explants (Kuang et al, 1998;Mizukami et al, 2004), Ulva lactuca protoplasts (Huang et al, 1996) and Gracilaria changii explants (Gan et al, 2003). In many of these cases, transient gene expression was achieved using the Cauliflower mosaic virus (CaMV) 35S promoter.…”

Section: Introductionmentioning

confidence: 99%

“…Transient expression of the lacZ gene with the SV40 promoter has also been observed in G. changii (Gan et al, 2003) and Haematococcus pluvialis (Teng et al, 2002). Recently, the use of endogenous promoters, targeted homologous recombination and matrix attachment regions (MARs) to increase reporter gene expression in P. yezoenisis has received much attention (Fukuda et al, 2008;Gong et al, 2007;Liu et al, 2003;Mizukami et al, 2004;Zuo et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Transformation of lacZ using different promoters in the commercially important red alga, Gracilaria gracilis

Huddy¹,

Meyers²,

Coyne³

2012

Afr. J. Biotechnol.

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“…In addition to gene delivery, another important factor for transgenesis is the selection of appropriate promoters. Although promoters for the chloroplast ribulose 1, 5-bisphosphate carboxylase/oxygenase (Rubisco) gene (Mizukami et al 2004), the beta-tubulin gene (Gong et al 2007) and the fucoxanthin chlorophyll a/c-binding protein (FCP) gene (Li et al 2009), as well as the much used CaMV35S promoter are available, the SV40 promoter has shown the most efficient capability for driving uniform exogenous gene expression in seaweeds without tissue specificity (Jiang et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement

et al. 2010

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This study investigated the delivery of a SV40 promoter driving lacZ gene into cells of Kappaphycus alvarezii using particle bombardment. Thallus pieces 0.5-0.8 mm in diameter and 1 cm in length were prepared as gene recipients. Bombardment parameters of 450 psi (rupture pressures) 9 6 cm (particle travel distances), 650 psi 9 6 cm, 1,100 psi 9 6 cm and 1,100 psi 9 9 cm were used. A significant increase in transformation efficiency from about 33% under the rupture pressure of 450 psi to 87% at 650 psi was observed in transformed thalli. Most of the positive cells appeared in epidermal cells bombarded at 450 psi, whereas positive signals were seen in both epidermal and medullary cells at 650 psi. No positive transient expression was detected at a bombardment of 1,100 psi, or in negative or blank controls. For the conditions tested, the best parameter was obtained at 650 psi at a distance of 6 cm. Thus, the strategy of taking vegetative thalli as recipients, using particle bombardment, and combining this with micro-propagation, together with developing an in vivo selectable marker, is a viable way to produce stable transformants, to eliminate chimeric expression, and to achieve transgenic breeding in K. alvarezii.

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Reporter gene introduction and transient expression in protoplasts of Porphyra yezoensis

Cited by 18 publications

References 21 publications

Efficient gusA transient expression in Porphyra yezoensis protoplasts mediated by endogenous beta-tubulin flanking sequences

Efficient gusA transient expression in Porphyra yezoensis protoplasts mediated by endogenous beta-tubulin flanking sequences

Transformation of lacZ using different promoters in the commercially important red alga, Gracilaria gracilis

Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement

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