สืบค้นงานวิจัย
Optimisation of electroporation and lipofection protocols to derive the black tiger shrimp cell line (Penaeus monodon)
Thansa K. - ไม่ระบุหน่วยงาน
ชื่อเรื่อง (EN): Optimisation of electroporation and lipofection protocols to derive the black tiger shrimp cell line (Penaeus monodon)
ผู้แต่ง / หัวหน้าโครงการ (EN): Thansa K.
บทคัดย่อ (EN): To achieve in creating permanent shrimp cell lines, cellular arrest of primary cells in the culture is needed to be firstly solved. Considering the insertion of some markers affecting cellular proliferation into primary haemocytes in order to produce the black tiger shrimp cell line and the very low percent of transduced cells previously reported in penaeid shrimps, these paved us the way to set up suitable gene delivery protocols to increase percent of transduced cells in the shrimp as our primary aim. In this study, electroporation and lipofection were used to transfer construct plasmids (pLL3.7 plasmids containing CMV promoters and pGL-IE1-126(A)-EGFP plasmids carrying WSSV IE1 promoters) into primary haemocytes. As it was difficult to distinguish between cells expressing EGFP signal and auto-fluorescence of many dead cells occurred by electroporation during the first 72 h of experiment; so, only lipofection was managed to deliver plasmids into primary cells. Surprisingly, numbers of suspected proliferative cells were derived after electroporation with no insertion of immortalising markers. These cells survived in vitro for up to 45 days with high rate of cell viability, but the number of viable cells decreased throughout the experiment. In addition, these cells expressed genes and proteins closely related to hyaline cells determined using RT-PCR and western blot. For the lipofection experiment, no green fluorescence signal was detected in any primary cell introduced with these plasmids, suggesting that plasmids were not successfully inserted into cells. Also, a number of primary haemocytes had the apoptotic cell death characteristic within 5 days after lipofection. These possibly result from using inappropriate lipofection protocol and chemical substances. In summary, finding out suitable protocols to elevate the percent of transduced cells is still necessary. Additionally, continuous shrimp cell lines would be possibly established by transforming suspected proliferative cells derived from electroporation in this study. © 2018 Elsevier Ltd
บทคัดย่อ: ไม่พบข้อมูลจากหน่วยงานต้นทาง
ภาษา (EN): en
เอกสารแนบ (EN): https://www.scopus.com/inward/record.uri?eid=2-s2.0-85049844332&doi=10.1016%2fj.fsi.2018.07.030&partnerID=40&md5=d69f508c481d14082a7c525f4db3bc64
เผยแพร่โดย (EN): มหาวิทยาลัยมหิดล
คำสำคัญ (EN): Promoter Regions, Genetic
เจ้าของลิขสิทธิ์ (EN): มหาวิทยาลัยมหิดล
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Thansa K.. "Optimisation of electroporation and lipofection protocols to derive the black tiger shrimp cell line (Penaeus monodon)". ไม่ระบุหน่วยงาน. ม.ป.ป.

Thansa K.. "Optimisation of electroporation and lipofection protocols to derive the black tiger shrimp cell line (Penaeus monodon)". ไม่ระบุหน่วยงาน. ม.ป.ป.. Print.



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Optimisation of electroporation and lipofection protocols to derive the black tiger shrimp cell line (Penaeus monodon)
Thansa K.
มหาวิทยาลัยมหิดล
ไม่ระบุวันที่เผยแพร่
Needle biopsy of the hepatopancreas of the black tiger shrimp Penaeus monodon with Penaeus monodon densovirus detection A novel function of bursicon in stimulation of vitellogenin expression in black tiger shrimp, Penaeus monodon Alteration of egg activation by protease inhibitors in the black tiger shrimp, Penaeus monodon A novel gonad-specific Argonaute 4 serves as a defense against transposons in the black tiger shrimp Penaeus monodon Characterization of PmSpӓtzle 1 from the black tiger shrimp Peneaus monodon A formulated double-stranded RNA diet for reducing Penaeus monodon densovirus infection in black tiger shrimp (ภาษาอังกฤษ) Functional characterization of a novel C-type lectin gene in the black tiger shrimp, Penaeus monodon Effects of fatty acids enrichment on polychaetes to be used as a maturation feed for the male black tiger shrimp (Penaeus monodon) Two new anti-apoptotic proteins of white spot syndrome virus that bind to an effector caspase (PmCasp) of the giant tiger shrimp Penaeus (Penaeus) monodon Australian red claw crayfish (Cherax quadricarinatus) is susceptible to yellow head virus (YHV) infection and can transmit it to the black tiger shrimp (Penaeus monodon)
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