scholarly journals Isolation and Homology Analysis of Alanine Aminotransferase Gene of Barley, Foxtail Millet, Cucumber, and Tomato

2020 ◽  
Vol 16 (2) ◽  
pp. 59
Author(s):  
Atmitri Sisharmini ◽  
Aniversari Apriana ◽  
Tri Joko Santoso ◽  
Bambang Sapta Purwoko ◽  
Nurul Khumaida ◽  
...  
Keyword(s):  
Phylogenetic Tree ◽  
Alanine Aminotransferase ◽  
Foxtail Millet ◽  
Amino Acid Sequences ◽  
Nitrogen Use ◽  
Engineering Research ◽  
Group Iii ◽  
Group I ◽  
Use Efficiency ◽  
Clustal Omega

<p>Overexpression of <em>alanine aminotransferase<strong> </strong></em>(<em>AlaAT</em>) gene can improve nitrogen use efficiency (NUE) in plants. The previous isolated <em>AlaAT</em> genes cannot be freely applied to generate NUE plants due to IPR restriction. Therefore, isolation of the gene from targeted monocot and dicot plants is necessary. The objectives of this study were to isolate <em>AlaAT</em> genes from barley, foxtail millet, cucumber, and tomato and analyze their homology to other isolated <em>AlaAT</em> genes in sequence databases (gene bank). Total RNA was isolated from roots of barley, foxtail millet, cucumber, and tomato, and then converted into cDNA using reverse transcription method. The cDNA was then cloned into <em>pGEM®-T Easy</em> plasmid and the verified clones were sequenced. The amino acid sequences were analyzed for their homologies using Clustal Omega software and phylogenetic tree was constructed. The results showed that the amino acids of <em>AlaAT</em> gene from barley was different from <em>AlaAT</em> genes of tomato and cucumber with homology level less than 80%. Phylogenetic tree predicted that <em>AlaAT</em> genes clustered into three groups with <em>AlaAT</em> genes of foxtail millet and barley clustered in one group together with other monocots in group I. <em>AlaAT</em> genes derived from dicots clustered into two groups<em> </em>in which<em> AlaAT</em> gene of tomato clustered in group II, while that of cucumber was in group III. The identity differences of <em>AlaAT</em> gene of tomato and that of cucumber as well as the estimates of the same enzymatic functions can open up enormous opportunities in genetic engineering research for the development of NUE rice.</p>

scholarly journals H13 Subtype Avian Influenza Viruses Are Crossing Host Species Barrier From Wild Waterfowl to Land Fowl

2021 ◽  
Author(s):  
Weiyang SUN ◽  
Menglin ZHAO ◽  
Zhijun YU ◽  
Yuanguo LI ◽  
Xinghai ZHANG ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Chicken Embryo Fibroblast ◽  
Amino Acid Sequences ◽  
Influenza Viruses ◽  
Post Inoculation ◽  
Species Barrier ◽  
Group Iii ◽  
Mammalian Cell Lines ◽  
Group I ◽  
Wild Waterfowl

Abstract The avian influenza virus H13 subtype circulates primarily in waterfowl. To explore the ability of the H13 virus to cross the host barriers, we genetically analysed two H13 isolates from wild birds in China and evaluated the infectivity of these subtypes in chickens. Genetic and molecular analyses showed differences in the lineages and amino acid sequences between the two subtypes; A/mallard/Dalian/DZ-137/2013 (H13N6) belonged to Group I, while A/Eurasian Curlew/Liaoning/ZH-385/2014 (H13N8) belonged to Group III. The nucleotide sequence results showed high homology (approximately 96.9%-100%) to sequences in GenBank. Neither H13 isolate replicated in adult chickens or 20-day-old chicks; however, the H13N8 strain replicated in 1- and 10-day-old chicks. Viruses were recovered from the nasal turbinate, tracheal, lung and colon tissues of chicks at 1, 3 and 5 days post-inoculation. The H13N6 isolates replicated inefficiently in 1-day-old chicks and did not replicate in 10-day-old chicks. Serological surveillance results showed that domestic chickens had a 4.6%-10.4% (15/328-34/328) positive antibody titre to the H13 virus. H13N6 and H13N8 isolates replicated in mammalian cell lines, including 293T, Madin-Darby canine kidney and chicken embryo fibroblast cells. Our results suggest that the AIV H13 subtype may cross the host barrier from wild waterfowl to land fowl.

Physiological analysis of nitrogen-efficient rice overexpressing alanine aminotransferase under different N regimes

Botany ◽  
2013 ◽  
Vol 91 (12) ◽  
pp. 866-883 ◽  
Author(s):  
Perrin H. Beatty ◽  
Rebecka T. Carroll ◽  
Ashok K. Shrawat ◽  
David Guevara ◽  
Allen G. Good
Keyword(s):  
Transgenic Rice ◽  
Nitrogen Use Efficiency ◽  
Alanine Aminotransferase ◽  
Nitrogen Use ◽  
Physiological Analysis ◽  
Use Efficiency ◽  
Amino Acid Levels ◽  
Transcriptome Response ◽  
Low Nitrogen ◽  
N Use

Cereal crop plants have low nitrogen (N) use efficiency, taking up only 30% to 50% of the applied N fertilizers, with the rest having the potential for loss into the environment as N pollution. One way to address this problem is to improve the nitrogen use efficiency of cereal crops using a transgenic approach. We developed alanine aminotransferase overexpressing rice, and we have previously determined that this modification provided an improved nitrogen-use phenotype to the engineered plants. In this study, the transgenic rice were grown in low, medium, and high nitrogen supply, and morphology, plant N levels, enzymatic activity, metabolite levels, and transcriptome response in the roots and shoots at active and maximum tillering at each N level were measured. The transcriptome response was analysed further using MapMan and PageMan to view multiple comparisons. The transgenic rice plants showed improved nitrogen use efficiency at medium and high N supply, but with few significant changes to the amino acid levels or to the transcriptome. The transgenic plants grown in high N showed up-regulation of transcripts associated with photosynthesis, non-melavonate pathway secondary metabolites, protein degradation, and many unknown function transcripts.

scholarly journals Laparoscopy hepatic biopsy through cauterization

2003 ◽  
Vol 33 (4) ◽  
pp. 703-707 ◽  
Author(s):  
Alexandra Pinheiro Fantinatti ◽  
Carlos Roberto Daleck ◽  
Newton Nunes ◽  
Antonio Carlos Alessi ◽  
João Moreira da Costa Neto ◽  
...  
Keyword(s):  
Alanine Aminotransferase ◽  
Hepatic Function ◽  
Radiofrequency Energy ◽  
Hepatic Biopsy ◽  
Group Iii ◽  
Applied Method ◽  
Group I ◽  
Healthy Dogs ◽  
Group Ii

Hepatic biopsy was realized through laparoscopy with simultaneous cauterization in the present study, whose principal aim was to evaluate the efficacy of the applied method and to study its effects in the liver of healthy dogs. Furthermore, we tried to verify the main hematological and chemistry profile alterations related to the hepatic function, and to investigate the viability of the fragments collected by histopathology. To attain this objective, 21 clinically healthy dogs, weighing between 10 and 15kg were submitted to hepatic biopsy with forceps connected to the cautery. Cautery was performed by applying radiofrequency energy at 45 watts. Forty-two hepatic biopsies through laparoscopy were conducted in the animals. At group I one hepatic fragment per animal was collected, at group II two hepatic fragments per animal were collected, and at group III three fragments were collected. Hematocrit and alanine-aminotransferase measurements were employed to evaluate the animals at the pre-operative period, at four and six hour post-operative, and at day 30 post-operative. The results revealed that the procedure was safe and effective for hepatic biopsy in dogs. There were no clinical alterations related to the technique. The fragments collected were viable for histopathology. Hepatic biopsy through laparoscopy with simultaneous cauterization is an effective and usefull method in dogs.

Engineering nitrogen use efficiency with alanine aminotransferase

2007 ◽  
Vol 85 (3) ◽  
pp. 252-262 ◽  
Author(s):  
Allen G. Good ◽  
Susan J. Johnson ◽  
Mary De Pauw ◽  
Rebecka T. Carroll ◽  
Nic Savidov ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Seed Yield ◽  
Nitrogen Use Efficiency ◽  
Nitrogen Fertilizer ◽  
Alanine Aminotransferase ◽  
Crop Productivity ◽  
Wild Type ◽  
Nitrogen Use ◽  
Use Efficiency ◽  
N Use

Nitrogen (N) is the most important factor limiting crop productivity worldwide. The ability of plants to acquire N from applied fertilizers is one of the critical steps limiting the efficient use of nitrogen. To improve N use efficiency, genetically modified plants that overexpress alanine aminotransferase (AlaAT) were engineered by introducing a barley AlaAT cDNA driven by a canola root specific promoter (btg26). Compared with wild-type canola, transgenic plants had increased biomass and seed yield both in the laboratory and field under low N conditions, whereas no differences were observed under high N. The transgenics also had increased nitrate influx. These changes resulted in a 40% decrease in the amount of applied nitrogen fertilizer required under field conditions to achieve yields equivalent to wild-type plants.

scholarly journals Purification, characterization and cDNA cloning of a phospholipase A2 inhibitor from the serum of the non-venomous snake Elaphe quadrivirgata

1999 ◽  
Vol 341 (1) ◽  
pp. 165-171 ◽  
Author(s):  
Kohji OKUMURA ◽  
Kuniharu MASUI ◽  
Seiji INOUE ◽  
Kiyoshi IKEDA ◽  
Kyozo HAYASHI
Keyword(s):  
Amino Acid ◽  
Phospholipase A2 ◽  
Gel Filtration ◽  
Protective Role ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Venomous Snake ◽  
Group Iii ◽  
Venomous Snakes ◽  
Group I

The serum of a non-venomous striated snake, Elaphequadrivirgata, was found to contain phospholipase A2 (PLA2) inhibitory proteins (PLIs). One of these inhibitors was purified by Sephadex G-200 gel filtration, Q-Sepharose FF ion-exchange chromatography and Butyl Sepharose 4FF hydrophobic chromatography. The purified PLI inhibited the enzymic activities of all PLA2 groups, including Elapidae venom (group-I), Viperidae venom (group-II) and honeybee PLA2s (group-III). The inhibitor was a 130 kDa glycoprotein consisting of two distinct subunits, A and B, of 30 and 29 kDa respectively; each of which was glycosylated with N-linked oligosaccharide chains. The cDNAs encoding the respective inhibitor subunits were isolated from a liver cDNA library by the use of probes, prepared by PCR, based on the partially determined amino-acid sequences of the corresponding subunits. The respective nucleotide sequences encoded 19-amino-acid-residue signal sequences, followed by 183- and 181-residue protein sequences for the A and B subunits respectively. The amino-acid sequences revealed that the E.quadrivirgata inhibitor corresponded to PLIγ, one of three kinds of inhibitors purified from venomous snakes. The existence of PLIγ in the serum of this non-venomous snake suggested that, besides having a protective role against the venom PLA2s of other venomous snakes, PLIγ has other important physiological functions in regulating local PLA2 activities; and thus it raises the possibility that PLIγ occurs in other animals, including mammals.

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