Production and characterization of fungal β-glucosidase and bacterial cellulases by tobacco chloroplast transformation

AbstractMedium-chain-length polyhydroxyalkanoates (mcl-PHAs) belong to the group of microbial polyesters. The key enzyme for mcl-PHA biosynthesis is type II PHA synthase. The genephaC2 encoding type II PHA synthase was placed under the control ofpsbA-pro andpsbA-ter of rice (Oryza sativa) to construct aphaC2 cassette, which was ligated with the screening marker geneaadAcassette (prrn–aadA–TpsbA-ter). These recombined fragments were cloned between the plastidrbcLandaccDgenes for targeting to the large single copy region of the chloroplast genome. A chloroplast transformation vector, pTC2, was constructed and introduced into the tobacco (Nicotiana tobacum) chloroplast genome by particle bombardment. PCR and Southern blot analysis confirmed stable integration ofphaC2 into the chloroplast genomes of T0and T1transgenic plants, and T1transgenic plants exhibited homoplasmy. The expression ofphaC2 at transcription level was detected by reverse transcriptase–polymerase chain reaction (RT-PCR). Recombinant transgenes in the tobacco chloroplast genome were maternally inherited and were not transmitted via pollen when out-crossed with untransformed female plants. To our knowledge, this is the first report on the stable transformation ofphaC2 encoding type II PHA synthase in tobacco via chloroplast genetic engineering.

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Characterization of replication origins flanking the 23S rRNA gene in tobacco chloroplast DNA

Plant Molecular Biology ◽

10.1007/bf00020210 ◽

1996 ◽

Vol 32 (4) ◽

pp. 693-706 ◽

Cited By ~ 19

Author(s):

Zhun Lu ◽

Muthusamy Kunnimalaiyaan ◽

Brent L. Nielsen

Keyword(s):

Chloroplast Dna ◽

23S Rrna ◽

Rrna Gene ◽

Replication Origins ◽

Tobacco Chloroplast ◽

23S Rrna Gene

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Molecular Characterization of atpA and atpB Deletion Mutants Induced in Chlamydomonas Reinhardth CW15 by Chloroplast Transformation

Photosynthesis: from Light to Biosphere ◽

10.1007/978-94-009-0173-5_499 ◽

1995 ◽

pp. 2119-2122

Author(s):

S. Leu ◽

J. Schlesinger ◽

Hu Dongli ◽

H. R. Fiedler ◽

H. Strotmann ◽

...

Keyword(s):

Molecular Characterization ◽

Chloroplast Transformation ◽

Deletion Mutants

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Characterization of Ribonucleoproteins from the Tobacco Chloroplast

Photosynthesis: Mechanisms and Effects ◽

10.1007/978-94-011-3953-3_693 ◽

1998 ◽

pp. 2957-2958 ◽

Cited By ~ 1

Author(s):

Takahiro Nakamura ◽

Masaru Ohta ◽

Mamoru Sugita ◽

Masahiro Sugiura

Keyword(s):

Tobacco Chloroplast

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Morphological Characterization of Mycobacteriophage R1

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051281 ◽

1974 ◽

Vol 32 ◽

pp. 254-255

Author(s):

B. L. Soloff ◽

T. A. Rado

Keyword(s):

Carbon Source ◽

Stationary Phase ◽

Growth Phase ◽

Minimal Medium ◽

Morphological Characterization ◽

Logarithmic Growth Phase ◽

Complete Lysis ◽

Lysogenic Culture ◽

Logarithmic Growth

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.

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AEM analysis of crystallization in Ti-based amorphous alloys

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100075142 ◽

1983 ◽

Vol 41 ◽

pp. 270-271

Author(s):

A.R. Pelton ◽

A.F. Marshall ◽

Y.S. Lee

Keyword(s):

Magnetic Properties ◽

Amorphous Alloys ◽

Amorphous Materials ◽

Isothermal Annealing ◽

Amorphous Matrix ◽

Nucleation And Growth ◽

Current Interest ◽

Amorphous Films ◽

Electrical And Magnetic Properties

Amorphous materials are of current interest due to their desirable mechanical, electrical and magnetic properties. Furthermore, crystallizing amorphous alloys provides an avenue for discerning sequential and competitive phases thus allowing access to otherwise inaccessible crystalline structures. Previous studies have shown the benefits of using AEM to determine crystal structures and compositions of partially crystallized alloys. The present paper will discuss the AEM characterization of crystallized Cu-Ti and Ni-Ti amorphous films.Cu60Ti40: The amorphous alloy Cu60Ti40, when continuously heated, forms a simple intermediate, macrocrystalline phase which then transforms to the ordered, equilibrium Cu3Ti2 phase. However, contrary to what one would expect from kinetic considerations, isothermal annealing below the isochronal crystallization temperature results in direct nucleation and growth of Cu3Ti2 from the amorphous matrix.

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Characterization of Coherent, Ordered Precipitates in Nickel-Base Alloys

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100071168 ◽

1973 ◽

Vol 31 ◽

pp. 144-145

Author(s):

B. H. Kear ◽

J. M. Oblak

Keyword(s):

Stable Phase ◽

Nickel Base Superalloy ◽

Alloy 718 ◽

Commercial Alloy ◽

Precipitate Morphology ◽

Continuous Precipitation ◽

Nickel Base ◽

Base Superalloy ◽

Strengthening Precipitate

A nickel-base superalloy is essentially a Ni/Cr solid solution hardened by additions of Al (Ti, Nb, etc.) to precipitate a coherent, ordered phase. In most commercial alloy systems, e.g. B-1900, IN-100 and Mar-M200, the stable precipitate is Ni3 (Al,Ti) γ′, with an LI2structure. In A lloy 901 the normal precipitate is metastable Nis Ti3 γ′ ; the stable phase is a hexagonal Do2 4 structure. In Alloy 718 the strengthening precipitate is metastable γ″, which has a body-centered tetragonal D022 structure.Precipitate MorphologyIn most systems the ordered γ′ phase forms by a continuous precipitation re-action, which gives rise to a uniform intragranular dispersion of precipitate particles. For zero γ/γ′ misfit, the γ′ precipitates assume a spheroidal.

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The Use of Advanced Analytical Techniques for Characterization of the Chemical, Physical, and Crystallographic Nature of a Surface

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100071107 ◽

1973 ◽

Vol 31 ◽

pp. 132-133 ◽

Cited By ~ 1

Author(s):

R. E. Herfert

Keyword(s):

Surface Characterization ◽

Analytical Techniques ◽

X Ray Diffraction ◽

Depth Of Penetration ◽

X Ray ◽

The Past ◽

Transmission Electron ◽

Scanning Electron

Studies of the nature of a surface, either metallic or nonmetallic, in the past, have been limited to the instrumentation available for these measurements. In the past, optical microscopy, replica transmission electron microscopy, electron or X-ray diffraction and optical or X-ray spectroscopy have provided the means of surface characterization. Actually, some of these techniques are not purely surface; the depth of penetration may be a few thousands of an inch. Within the last five years, instrumentation has been made available which now makes it practical for use to study the outer few 100A of layers and characterize it completely from a chemical, physical, and crystallographic standpoint. The scanning electron microscope (SEM) provides a means of viewing the surface of a material in situ to magnifications as high as 250,000X.

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Skeletal elements of crinoid echinoderms: High-resolution structural and microanalytical results

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100074768 ◽

1983 ◽

Vol 41 ◽

pp. 194-195

Author(s):

D. F. Blake ◽

L. F. Allard ◽

D. R. Peacor

Keyword(s):

High Resolution ◽

Marine Invertebrates ◽

Sea Urchins ◽

Structural Features ◽

Sea Stars ◽

High Resolution Tem ◽

Relationship Of ◽

The Relationship ◽

Insight Into

Echinodermata is a phylum of marine invertebrates which has been extant since Cambrian time (c.a. 500 m.y. before the present). Modern examples of echinoderms include sea urchins, sea stars, and sea lilies (crinoids). The endoskeletons of echinoderms are composed of plates or ossicles (Fig. 1) which are with few exceptions, porous, single crystals of high-magnesian calcite. Despite their single crystal nature, fracture surfaces do not exhibit the near-perfect {10.4} cleavage characteristic of inorganic calcite. This paradoxical mix of biogenic and inorganic features has prompted much recent work on echinoderm skeletal crystallography. Furthermore, fossil echinoderm hard parts comprise a volumetrically significant portion of some marine limestones sequences. The ultrastructural and microchemical characterization of modern skeletal material should lend insight into: 1). The nature of the biogenic processes involved, for example, the relationship of Mg heterogeneity to morphological and structural features in modern echinoderm material, and 2). The nature of the diagenetic changes undergone by their ancient, fossilized counterparts. In this study, high resolution TEM (HRTEM), high voltage TEM (HVTEM), and STEM microanalysis are used to characterize tha ultrastructural and microchemical composition of skeletal elements of the modern crinoid Neocrinus blakei.

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