Search results for "EGFP"
showing 6 items of 6 documents
Bromoviridae dzimtas augu vīrusu apvalka proteīnu N-terminālo daļu īpašību pētījumi
2016
Bakalaura darba mērķis ir pārbaudīt ekspresēto un attīrīto ApMV un CMV CP N-gala sajūgtos domēnus ar eGFP dimerizācijas-oligomerizācijas īpašības, kas varētu turpmāk tikt izmantotas imunizācijas eksperimentiem. Darba gaitā tika izveidotas trīs konstrukcijas, kur CP N-terminālie domēni tika ģenētiski savienoti ar eGFP modeļproteīnu un sešu histidīnu atkārtojumiem sajūgto proteīnu attīrīšanai. Sajūgtie proteīni tika noekspresēti un, attīrīti ar afīno hromatogrāfiju. Nukleīnskābes kustīguma nobīdes eksperimenti parādīja, ka ar Zn2+ audzētam nApMV-eGFP ir augstāka afinitāte pret CP mRNS un PCR, nekā audzētam bez Zn2+. Visām N-gala proteīniem ir augstāka afinitāte pret CP mRNS nekā pret CP PCR. …
RNA Transfer by Electroporation into Mature Dendritic Cells Leading to Reactivation of Effector-Memory Cytotoxic T Lymphocytes: A Quantitative Analys…
2005
Previous studies have analyzed transfer of RNA-encoded tumor-associated antigens (TAAs) into immature dendritic cells (DCs) because of their exceptional ability to internalize antigens. Concerns have been raised regarding the use of immature DCs in clinical studies because of their capacity to tolerize T cells. Therefore, we focused on optimizing RNA transfer into mature DCs using the method of electroporation and obtained high protein expression in 90% of mature DCs. Particular emphasis was placed on quantifying RNA transfer. Reconstitution of peptide-MHC (pMHC) ligands on RNA-pulsed DCs was measured with the help of effector-memory cytotoxic T lymphocytes (CTLs) specific for the melanoma-…
EGFP Reporters for Direct and Sensitive Detection of Mutagenic Bypass of DNA Lesions
2020
The sustainment of replication and transcription of damaged DNA is essential for cell survival under genotoxic stress
Determination of a conformational change in filamin A with Förster resonance energy transfer
2011
Filamins are large rod-like proteins that cross-link actin filaments into three-dimensional networks. They also bind to a plethora of proteins with distinct functions showing that they have a versatile role in cells. Functional filamins are dimers consisting of an N-terminal actin binding domain followed by 24 immunoglobulin-like domains. The most C-terminal domains mediate the dimerization. Two hinge regions are located between the domains 15 and 16 and 23 and 24, respectively, and produce structural flexibility that is essential for the protein function. The domains 18-19 and 20-21 are folded in a pairwise manner in which the first β strands of the even numbered domains are folded along w…
Systematic Study of a Library of PDMAEMA-Based, Superparamagnetic Nano-Stars for the Transfection of CHO-K1 Cells.
2017
The introduction of the DNA into mammalian cells remains a challenge in gene delivery, particularly in vivo. Viral vectors are unmatched in their efficiency for gene delivery, but may trigger immune responses and cause severe side-reactions. Non-viral vectors are much less efficient. Recently, our group has suggested that a star-shaped structure improves and even transforms the gene delivery capability of synthetic polycations. In this contribution, this effect was systematically studied using a library of highly homogeneous, paramagnetic nano-star polycations with varied arm lengths and grafting densities. Gene delivery was conducted in CHO-K1 cells, using a plasmid encoding a green fluore…
Cilvēka nātrija tauroholāta ko-transportējoša polipeptīda klonēšana un ekspresija zīdītāju šūnās
2015
Neskatoties uz to, ka ir izstrādāta droša un efektīva hepatīta B vakcīna, kā arī vesela rinda pret hepatīta B vīrusu (HBV) vērstu ķīmisku līdzekļu, pasaulē ir >350 miljonu hronisko HBV nēsātāju, no kuriem 1 miljons mirst katru gadu. Galvenais jaunu terapijas veidu izstrādāšanas kavēklis ir efektīvas HBV infekciju un replikāciju nodrošinošas in vitro modeļsistēmas trūkums. Pateicoties revolucionāram atklājumam par NTCP (sodium taurocholate cotransporting peptide) lomu HBV infekcijas procesā pašlaik tiek aktīvi izstrādātās jaunās HBV in vitro infekcijas modeļsistēmas. Mūsu darbā NTCP receptoru ekspresējošo šūnu iegūšanai tika izmantota efektīva Semliki Meža vīrusa (SFV) replikona ekspresijas …