Cloning and expression of cDNAs encoding α1,3-fucosyltransferase homologues from Arabidopsis thaliana1The cDNA sequences referred to in this publication have been deposited with the EMBL database under the numbers AJ404860 (FucTA), AJ404861 (FucTB) and AJ404862 (FucTC).1

Global warming has gradually reduced the control efficacy of Agasicles hygrophila against the invasive weed Alternanthera philoxeroides. To better understand the summer collapse of A. hygrophila populations, we cloned the cDNA sequences of the high temperature-sensing TRPA1, Painless, and Pyrexia from A. hygrophila, and analyzed their temporal expressions and the impacts of high temperatures on their expression in eggs, the most vulnerable stage of A. hygrophila to hot temperatures. All the three genes obtained had the signature domains of TRPA channels and were constitutively expressed in eggs, larvae (L1, L2, L3), pupae, and adults, but AhPainless had the highest expression, followed by AhPyrexia, and AhTRPA1. The lowest and highest expression stages were adult and pupae for AhTRPA1, egg and L3 for AhPainless, and pupae/adult and L2 for AhPyrexia. The expressions of AhTRPA1, AhPainless, and AhPyrexia remained low at the preferred temperature range of 25–28 °C, elevated to their peak levels at 37.5, 30, and 30 °C, respectively, then fell to their 25–28 °C levels (AhTRPA1, AhPainless) or a lower level (AhPyrexia) at one or more temperatures >30 or 37.5 °C. These results suggest that their temperature-sensing roles and importance may be different, stage-specific, and their expression may be decoupled from their activation.

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Cloning and expression analysis of full length mouse cDNA sequences encoding the transformation associated protein p53

Nucleic Acids Research ◽

10.1093/nar/12.14.5609 ◽

1984 ◽

Vol 12 (14) ◽

pp. 5609-5626 ◽

Cited By ~ 41

Author(s):

John R. Jenkins ◽

Karran Rudge ◽

Shelagh Redmond ◽

Alison Wade-Evans

Keyword(s):

Expression Analysis ◽

Full Length ◽

Cloning And Expression ◽

Cdna Sequences ◽

Mouse Cdna ◽

Protein P53

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The full-length cDNA cloning and expression profiles of 14-3-3 genes from the mud crab Scylla paramamosain Estampador, 1949

Crustaceana ◽

10.1163/15685403-00003480 ◽

2015 ◽

Vol 88 (10-11) ◽

pp. 1065-1078 ◽

Cited By ~ 4

Author(s):

Miao-An Shu ◽

Chen Long ◽

Wei-Ren Dong ◽

Pan Zhang ◽

Bin-Peng Xu ◽

...

Keyword(s):

Expression Profiles ◽

Expression Patterns ◽

Full Length ◽

Cloning And Expression ◽

Scylla Paramamosain ◽

Mud Crab ◽

Phylogenetic Tree Analysis ◽

Specific Expression ◽

Full Length Cdna ◽

Cdna Sequences

Two full-length cDNA sequences of 14-3-3 genes were isolated from the mud crab, Scylla paramamosain Estampador, 1949, using RT-PCR and RACE (rapid-amplification of cDNA ends). Sequence analysis indicates that both 14-3-3 genes contain an open reading frame of 744 bp with a deduced 247-amino-acid protein. The gene and protein sequences of Sp14-3-3 genes show 96 and 97% identity, respectively, and both Sp14-3-3 cluster together with other animal 14-3-3 proteins in phylogenetic tree analysis. Tissue specific expression analysis reveals that both Sp14-3-3 genes are ubiquitously expressed, however, their expression patterns are different from each other. Expressions of both Sp14-3-3 genes are sensitive to salinity decrease, even a 5 ppt drop from 30 ppt to 25 ppt, indicating that Sp14-3-3 genes are involved in osmoregulation. Our observations may contribute to a better understanding of the molecular and functional evolution of the 14-3-3 family in both crustaceans and (in)vertebrates as a whole.

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cDNA cloning and expression of a gamma-aminobutyric acidA receptor epsilon-subunit in rat brain

European Journal of Neuroscience ◽

10.1046/j.1460-9568.2000.01343.x ◽

2000 ◽

Vol 12 (12) ◽

pp. 4318-4330 ◽

Cited By ~ 7

Author(s):

Nathalie Moragues ◽

Philippe Ciofi ◽

Pierrette Lafon ◽

Marie-Francoise Odessa ◽

Gerard Tramu ◽

...

Keyword(s):

Rat Brain ◽

Cdna Cloning ◽

Cloning And Expression ◽

Epsilon Subunit

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Cloning and Expression of Recombinant Human Insulin Gene in Pichia pastoris

International Journal of Pharmaceutical Quality Assurance ◽

10.25258/ijpqa.10.1.22 ◽

2019 ◽

Vol 10 (01) ◽

Author(s):

Rafid A. Abdulkareem

Keyword(s):

Pichia Pastoris ◽

Human Insulin ◽

Expression Vector ◽

Expression System ◽

Insulin Gene ◽

Cloning And Expression ◽

Pcr Analysis ◽

Major Band ◽

Human Insulin Gene ◽

Pichia Pastoris Expression System

The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).

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