Real-time quantitative PCR for evaluating murine thymic function / 南方医科大学学报
Journal of Southern Medical University
; (12): 62-65, 2006.
Article
en Zh
| WPRIM
| ID: wpr-234195
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To establish a real-time quantitative PCR method for detecting the levels of the signal joint T cell receptor excision circles (sjTRECs) in murine thymocytes and spleen lymphocytes for determining the amount of naive T cells and evaluating the thymic function.</p><p><b>METHODS</b>The genomic DNA was extracted from murine thymocytes and splenocytes for PCR amplification of the target fragments. After purification of the PCR product, the recombination-activating gene 2 (RAG(2)) fragment was cloned into pGEMT-Easy vector to construct the standard plasmid. After PCR optimization, the standard curve was obtained and the samples (thymocytes and splenocytes of BALB/c and C(57)BL/6 mice) were detected for sjTRECs by real-time quantitative PCR.</p><p><b>RESULTS</b>The standard plasmid was correctly constructed, and the standard curve with high reliability was obtained. No statistical difference was observed in sjTREC contents in the T lymphocytes between the two mouse strains.</p><p><b>CONCLUSIONS</b>Real-time quantitative PCR for sjTREC analysis is established successfully, which offers an important means for thymic function analysis and a reliable model establishment for study the thymus.</p>
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Timo
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Receptores de Antígenos de Linfocitos T
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Linfocitos T
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Reordenamiento Génico de Linfocito T
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Reacción en Cadena de la Polimerasa
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Recuento de Linfocitos
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Biología Celular
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Alergia e Inmunología
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Genética
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Metabolismo
Límite:
Animals
Idioma:
Zh
Revista:
Journal of Southern Medical University
Año:
2006
Tipo del documento:
Article