Identification, characterization and quantitative analysis of NAD-malate dehydrogenase from the marine rhodophyte Pyropia haitanensis

2015 ◽  
Vol 58 (4) ◽  
Author(s):  
Bao Yu Zhang ◽  
Zhao Jun Hou ◽  
Guang Ce Wang ◽  
Guang Peng

AbstractMalate dehydrogenase (MDH) is an enzyme that catalyzes the interconversion of malate and oxaloacetate substrates and is widely distributed from prokaryotes to eukaryotes. It plays crucial roles in many important metabolic pathways and includes different isoforms based on coenzyme specificity and cellular localization. To study MDH in rhodophytes, we obtained a full-length cDNA clone (here designated

2021 ◽  
Vol 166 (5) ◽  
pp. 1427-1431
Author(s):  
Buyang Chen ◽  
Qi Lin ◽  
Yueyan Yin ◽  
Liangliang Jiang ◽  
Fang Wang ◽  
...  

2008 ◽  
Vol 74 (6) ◽  
pp. 434-437 ◽  
Author(s):  
Atsushi Ohkawa ◽  
Noriko Ishikawa-Suehiro ◽  
Seiichi Okuda ◽  
Tomohide Natsuaki

1987 ◽  
Vol 15 (16) ◽  
pp. 6740-6740 ◽  
Author(s):  
Hans-Jürgen Schmelzer ◽  
Gerhard Gross ◽  
Georg Widera ◽  
Hubert Mayer

1994 ◽  
Vol 5 (12) ◽  
pp. 1301-1310 ◽  
Author(s):  
S W Clark ◽  
O Staub ◽  
I B Clark ◽  
E L Holzbaur ◽  
B M Paschal ◽  
...  

An examination of human-expressed sequence tags indicated the existence of an isoform of centractin, an actin-related protein localized to microtubule-associated structures. Using one of these tags, we isolated and determined the nucleotide sequence of a full-length cDNA clone. The protein encoded represents the first example of multiple isoforms of an actin-related protein in a single organism. Northern analysis using centractin-specific probes revealed three species of mRNA in HeLa cells that could encode centractin isoforms. One mRNA encodes the previously-identified centractin (now referred to as alpha-centractin). The full-length cDNA clone isolated using the expressed sequence tag encodes a new member of the centractin family, beta-centractin. A probe specific for alpha-centractin hybridized to the third species of mRNA observed (referred to as gamma-centractin). Comparisons of Northern blots of human tissues indicated that alpha-centractin and beta-centractin mRNAs are equally distributed in all populations of mRNA examined, whereas the expression of gamma-centractin appears to be tissue specific. The amino acid sequence of beta-centractin, deduced from the cDNA, indicates a 91% identity with alpha-centractin, increasing to 96% similarity when conservative amino acid changes are taken into account. As antibodies previously raised against alpha-centractin reacted only poorly with beta-centractin, new antibodies were produced and combined with two-dimensional gel electrophoresis to discriminate the two isoforms. Using this system, the subcellular distribution of the alpha- and beta-isoforms were determined. Both isoforms were found predominantly in the cytosolic fraction as a part of a previously identified 20S complex (referred to as the dynactin complex) with no evidence for a free pool of either isoform. The isoforms were found in a constant ratio of approximately 15:1 (alpha:beta) in the dynactin complex.


Virology ◽  
1995 ◽  
Vol 206 (1) ◽  
pp. 108-115 ◽  
Author(s):  
C. Brugidou ◽  
C. Holt ◽  
M. Ngon A Yassi ◽  
S. Zhang ◽  
R. Beachy ◽  
...  

2010 ◽  
Vol 142 (1-2) ◽  
pp. 3-12 ◽  
Author(s):  
Katrin Mischkale ◽  
Ilona Reimann ◽  
J. Zemke ◽  
P. König ◽  
Martin Beer

1998 ◽  
Vol 72 (1) ◽  
pp. 380-387 ◽  
Author(s):  
J. J. M. Meulenberg ◽  
J. N. A. Bos-de Ruijter ◽  
R. van de Graaf ◽  
G. Wensvoort ◽  
R. J. M. Moormann

ABSTRACT The 5′-terminal end of the genomic RNA of the Lelystad virus isolate (LV) of porcine reproductive and respiratory syndrome virus was determined. To construct full-length cDNA clones, the 5′-terminal sequence was ligated to cDNA clones covering the complete genome of LV. When RNA that was transcribed in vitro from these full-length cDNA clones was transfected into BHK-21 cells, infectious LV was produced and secreted. The virus was rescued by passage to porcine alveolar lung macrophages or CL2621 cells. When infectious transcripts were transfected to porcine alveolar lung macrophages or CL2621 cells, no infectious virus was produced due to the poor transfection efficiency of these cells. The growth properties of the viruses produced by BHK-21 cells transfected with infectious transcripts of LV cDNA resembled the growth properties of the parental virus from which the cDNA was derived. Two nucleotide changes leading to a unique PacI restriction site directly downstream of the ORF7 gene were introduced in the genome-length cDNA clone. The virus recovered from this mutated cDNA clone retained the PacI site, which confirmed the de novo generation of infectious LV from cloned cDNA. These results indicate that the infectious clone of LV enables us to mutagenize the viral genome at specific sites and that it will therefore be useful for detailed molecular characterization of the virus, as well as for the development of a safe and effective live vaccine for use in pigs.


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