Construction of fetal mesenchymal stem cell cDNA subtractive library / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 89-92, 2002.
Artigo
em Chinês
| WPRIM
| ID: wpr-258104
ABSTRACT
<p><b>UNLABELLED</b>To identify differentially expressed genes between fetal mesenchymal stem cell (MSC) and adult MSC, especially specified genes expressed in fetal MSC, a cDNA subtractive library of fetal MSC was constructed using suppression subtractive hybridization (SSH) technique. At first, total RNA was isolated from fetal and adult MSC. Using SMART PCR synthesis method, single-strand and double-strand cDNAs were synthesized. After Rsa I digestion, fetal MSC cDNAs were divided into two groups and ligated to adaptor 1 and adaptor 2 respectively. Results showed that the amplified library contains 890 clones. Analysis of 890 clones with PCR demonstrated that 768 clones were positive. The positive rate is 86.3%. The size of inserted fragments in these positive clones was between 0.2 - 1 kb, with an average of 400 - 600 bp.</p><p><b>CONCLUSION</b>SSH is a convenient and effective method for screening differentially expressed genes. The constructed cDNA subtractive library of fetal MSC cDNA lays solid foundation for screening and cloning new and specific function related genes of fetal MSC.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Células-Tronco
/
Desoxirribonucleases de Sítio Específico do Tipo II
/
Biblioteca Gênica
/
Reação em Cadeia da Polimerase
/
Clonagem Molecular
/
DNA Complementar
/
Biologia Celular
/
Feto
/
Genética
/
Mesoderma
Limite:
Humanos
Idioma:
Chinês
Revista:
Journal of Experimental Hematology
Ano de publicação:
2002
Tipo de documento:
Artigo
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