Regeneration of vgb-transgenic poplar (Populus alba×P. glandulosa) and the primary observation of growth

2006 ◽  
Vol 3 (1) ◽  
pp. 59-64 ◽  
Author(s):  
Zhang Bing-Yu ◽  
Su Xiao-Hua ◽  
Li Yi-Liang ◽  
Huang Qin-Jun ◽  
Zhang Xiang-Hua ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Blot Hybridization ◽  
Pcr Amplification ◽  
First Year ◽  
Populus Alba ◽  
Rt Pcr ◽  
Dot Blot ◽  
Transgenic Lines ◽  
Polymerase Chain ◽  
Morphological Differences

AbstractIncreasing the growth rate is especially important for low-quality wood applications, so this has become an important goal in poplar breeding. The present study describes the transfer of Vitreoscilla haemoglobin (VHb) gene (vgb) driven by constitutive promoters, by Agrobacterium tumefaciens into poplar (Populus alba×P. glandulosa). From about 450 leaf discs used for transformation, 60 Kan-resistant plants were obtained, and 52 proved to be true transgenic plants. The transgenic nature of these plants was confirmed by polymerase chain reaction (PCR) amplification and Southern dot blot hybridization. The expression of vgb gene in transgenic plants was confirmed by reverse transcriptase-PCR (RT-PCR). The performance of the transgenic lines was evaluated during the first year of growth in a greenhouse. These plants showed no significant stable morphological differences from the untransformed plants. Among them, three vgb-transgenic lines exhibited noticeably higher growth rates in terms of height and diameter.

Feasibility of Qualitative Testing of BCR-ABL and JAK2 V617F in Suspected Myeloproliperative Neoplasm (MPN) Using RT-PCR Reversed Dot Blot Hybridization (RT-PCR RDB)

2019 ◽  
Vol 19 (4) ◽  
pp. 220-227
Author(s):  
Najmiatul Masykura ◽  
Ummu Habibah ◽  
Siti Fatimah Selasih ◽  
Soegiarto Gani ◽  
Cosphiadi Irawan ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Jak2 V617f ◽  
Rt Pcr ◽  
Dot Blot ◽  
Dot Blot Hybridization

scholarly journals Altered Patterns of Gene Expression in Arabidopsis Elicited by Cauliflower Mosaic Virus (CaMV) Infection and by a CaMV Gene VI Transgene

1999 ◽  
Vol 12 (5) ◽  
pp. 377-384 ◽  
Author(s):  
Chiara Geri ◽  
Edi Cecchini ◽  
Maria E. Giannakou ◽  
Simon N. Covey ◽  
Joel J. Milner
Keyword(s):  
Gene Expression ◽  
Transgenic Plants ◽  
Mosaic Virus ◽  
Rna Binding ◽  
Important Determinant ◽  
Blot Hybridization ◽  
Cauliflower Mosaic Virus ◽  
Slot Blot Hybridization ◽  
Pcr Products ◽  
Polymerase Chain

Cauliflower mosaic virus (CaMV) gene VI protein (P6) is an important determinant of symptom expression. Differential display polymerase chain reaction (PCR) was used to identify changes in gene expression in Arabidopsis elicited by a P6 transgene that causes a symptomatic phenotype. We used slot blot hybridization to measure the abundance of mRNAs complementary to 66 candidate PCR products in transgenic, CaMV-infected, and uninfected Arabidopsis plants. CaMV-infected and P6 transgenic plants showed broadly similar changes in abundance of mRNA species. In P6 transgenic plants we detected 18 PCR products that showed unambiguous changes in abundance plus another 15 that showed more limited changes (approximately twofold). CaMV-infected plants showed 17 unambiguous and 13 limited changes. Down-regulated species include those encoding a novel, phenol-like sulfotransferase, and a glycine-rich, RNA-binding protein. Up-regulated species included ones encoding an myb protein, glycine-rich and stress-inducible proteins, and a member of a previously unreported gene family. CaMV infection causes alterations in expression of many Arabidopsis genes. Transgene-mediated expression of P6 mimics virus infection in its effect on host gene expression, providing a potential mechanism for this process.

scholarly journals A simple approach to prenatal diagnosis of beta-thalassemia in a geographic area where multiple mutations occur

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1357-1360 ◽  
Author(s):  
SP Cai ◽  
JZ Zhang ◽  
DH Huang ◽  
ZX Wang ◽  
YW Kan
Keyword(s):  
Southern China ◽  
Globin Gene ◽  
Blot Hybridization ◽  
Geographic Area ◽  
Simple Approach ◽  
Beta Thalassemia ◽  
Beta Globin ◽  
Dot Blot ◽  
Polymerase Chain

Abstract We describe a simple approach for detecting beta-thalassemia mutations in geographic areas such as southern China where multiple mutations are known to occur. Segments of the beta-globin gene were amplified in vitro by using the polymerase chain reaction. Dot blot hybridization of the amplified DNA with oligonucleotide probes corresponding to the six mutations found in southern China could directly identify the mutations causing beta-thalassemia in the affected families. The increased number of target sequences after amplification allows the use of 35S-labeled probes, which are reusable for up to 3 months. The mutations can be determined in two days.

scholarly journals Persistent Detection and Infectious Potential of SARS-CoV-2 Virus in Clinical Specimens from COVID-19 Patients

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1384
Author(s):  
Michael Zapor
Keyword(s):  
Pcr Amplification ◽  
World Health ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Distinctive Features ◽  
Clinical Specimens ◽  
Polymerase Chain ◽  
Health Organization ◽  
Infectious Potential ◽  
Infective Potential

The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) that emerged in December 2019 as the causative agent of Coronavirus 2019 (COVID-19) and was declared a pandemic by the World Health Organization in March 2020 has several distinctive features, including extensive multiorgan involvement with a robust systemic inflammatory response, significant associated morbidity and mortality, and prolonged persistence of viral RNA in the clinical specimens of infected individuals as detected by Reverse Transcription Polymerase Chain Reaction (RT-PCR) amplification. This review begins with an overview of SARS-CoV-2 morphology and replication and summarizes what is known to date about the detection of the virus in nasal, oropharyngeal, and fecal specimens of patients who have recovered from COVID-19, with a focus on the factors thought to contribute to prolonged detection. This review also provides a discussion on the infective potential of this material from asymptomatic, pre-symptomatic, and convalescing individuals, to include a discussion of the relative persistence and infectious potential of virus in clinical specimens recovered from pediatric COVID-19 patients.

scholarly journals Enhancing the expression of ARK1 genes in poplar leads to multiple branches and transcriptomic changes

2020 ◽  
Vol 7 (9) ◽  
pp. 201201
Author(s):  
Xiaozhen Liu ◽  
Zhiming Zhang ◽  
Wen Bian ◽  
Anan Duan ◽  
Hanyao Zhang
Keyword(s):  
Transgenic Plants ◽  
Quantitative Polymerase Chain Reaction ◽  
Plant Growth And Development ◽  
Expression Levels ◽  
Qpcr Analysis ◽  
Transgenic Lines ◽  
Polymerase Chain ◽  
Transcriptomic Changes ◽  
Insight Into ◽  
Poplar Seedlings

The ARBORKNOX1 ( ARK1 ) gene is an important gene for regulating plant growth and development; however, transcriptomic responses of enhancing expression of ARK1 gene in poplar are poorly investigated. To provide insight into the gene function of the ARK1 gene in poplar, the ARK1 transgenic poplar ‘717' and ‘84 K' lines were obtained, the morphology of transgenic plants was observed, and transcriptome profiles were compared. The results showed that there were multiple branches in ARK1 transgenic seedlings compared with non-transgenic seedlings. The results of transcriptome analysis showed that there were significant differences in transcriptome profiles between the transgenic lines of ‘717' and ‘84 K', and between non-transgenic lines (CK) and transgenic plants. The real-time quantitative polymerase chain reaction (RT-qPCR) analysis confirmed the expression levels of the genes involved in the pathway of zeatin biosynthesis and brassinosteroid biosynthesis. The increase in expression levels of AHP and CYCD3 was related to multiple branches. Enhancing the expression of the ARK1 gene in poplar seedlings leads to multiple branches and transcriptomic changes.

RT-PCR analysis of Na+/H+ exchanger mRNAs in rat medullary thick ascending limb

1995 ◽  
Vol 268 (6) ◽  
pp. F1224-F1228 ◽  
Author(s):  
P. Borensztein ◽  
M. Froissart ◽  
K. Laghmani ◽  
M. Bichara ◽  
M. Paillard
Keyword(s):  
Rat Kidney ◽  
Pcr Amplification ◽  
Restriction Enzymes ◽  
Pcr Analysis ◽  
Thick Ascending Limb ◽  
Rt Pcr ◽  
Medullary Thick Ascending Limb ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Nhe Isoforms

The thick ascending limb (TAL) of rat kidney absorbs bicarbonate secondary to proton secretion, but displays both basolateral and luminal Na+/H+ exchange (NHE) activity. Several NHE genes, including NHE-1, NHE-2, NHE-3, and NHE-4, are expressed in the kidney. To identify the NHE isoforms expressed in the rat medullary TAL (MTAL), we used the reverse transcription-polymerase chain reaction (RT-PCR) to detect the mRNAs for NHE in microdissected MTAL. RT-PCR amplification from total RNA was performed between two specific primers for each NHE isoform. In rat kidney homogenate, the four NHE isoform mRNAs were detected, and the identity of the PCR products was demonstrated by the sizes of the fragments, digestion with restriction enzymes, and Southern blot analysis. In microdissected rat MTAL, NHE-3 was strongly expressed and NHE-1 mRNA was also detected, whereas NHE-2 and NHE-4 mRNAs were not detected. Therefore, NHE-3 could be the apical Na+/H+ exchanger, and NHE-1 could be the basolateral isoform in the MTAL.

scholarly journals Distribution and Diversity of Geminiviruses in Trinidad and Tobago

1998 ◽  
Vol 88 (12) ◽  
pp. 1262-1268 ◽  
Author(s):  
Pathmanathan Umaharan ◽  
Malla Padidam ◽  
Ralph H. Phelps ◽  
Roger N. Beachy ◽  
Claude M. Fauquet
Keyword(s):  
Trinidad And Tobago ◽  
Sequence Similarity ◽  
Blot Hybridization ◽  
Pcr Amplification ◽  
Sweet Pepper ◽  
Yellow Mosaic Virus ◽  
Restriction Enzyme Digestion ◽  
Weed Species ◽  
Dot Blot ◽  
Degenerate Primers

Seven crop and eight weed species from 12 agricultural locations in Trinidad and Tobago were assayed for the presence of whitefly-transmitted geminiviruses (WTGs) by using dot blot hybridization and polymerase chain reaction (PCR) amplification of the N-terminal coat protein sequence with degenerate primers. The amplified fragments were cloned and analyzed by restriction enzyme digestion to determine fragment length polymorphism among the cloned fragments. Representative clones were then sequenced and subjected to phylogenetic analysis to determine the sequence similarity to known WTGs. WTGs were found in every location sampled and in 10 of the 15 species investigated: Lycopersicon esculentum(tomato), Capsicum annuum (pepper), Capsicum frutescens (sweet pepper), Abelmoschus esculentus (okra), Phaseolus vulgaris (beans), Alternanthera tenella, Desmodium frutescens, Euphorbia heterophylla, Malva alceifolia, and Sida acuta. The geminiviruses infecting these plants were closely related to potato yellow mosaic virus from Venezuela (PYMV-VE) and tomato leaf curl virus from Panama (ToLCV-PA). However, in pepper, sweet pepper, okra, Alternanthera tenella, Euphorbia heterophylla, Des-modium frutescens, and in one sample of tomato, a PYMV-VE-related virus was found in mixed infections with a virus related to pepper huasteco virus. Full-length infectious DNA-A and DNA-B of a tomato-infecting geminivirus from Trinidad and Tobago were cloned and sequenced. DNA-A appears to be a recombinant derived from PYMV-VE or ToLCV-PA, and Sida golden mosaic from Honduras. The implications of these findings in the control of WTGs are discussed.

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