Effect of α -1,2 fucosyltransferase gene 682A> G and 547_552delAG mutations on the activity of fucosyltransferase / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
; (6): 591-594, 2014.
Article
en Zh
| WPRIM
| ID: wpr-291723
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ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of α -1,2 fucosyltransferase (FUT1) gene 682A> G and 547_552delAG mutations on the expression of FUT1 mRNA and activity of α -1,2 fucosyltransferase.</p><p><b>METHODS</b>Recombinant expression vectors of FUT1 682A> G and FUT1 547_552delAG were constructed and transfected into COS-7 cells for stable expression screening. Expression of FUT1 mRNA was determined using real-time quantitative PCR. The activity of FUT1 was measured with high-performance liquid chromatography.</p><p><b>RESULTS</b>Stably transfected COS-7 cells with wild type FUT1, FUT1 682A> G and FUT1 547_552delAG were respectively obtained. The FUT1 mRNA level of transfected cells with 682A> G and 547_552delAG recombination vectors have measured 101.69% and 102.79% compared with that of wild type FUT1 transfected cells. A specific protein band with about 46 kD was confirmed in the 682A> G transfected cell lysates by SDS-PAGE electrophoresis and Western blotting with 6× His Tag antibody. Similar protein was not identified in the 547_552delAG cells lysates. Enzymes activity of FUT1 682A> G has measured 61.01% compared with wild type FUT1 protein, whilst the activity of FUT1 547_557delAG was completely abolished.</p><p><b>CONCLUSION</b>FUT1 682A> G and 547_552delAG mutations do not affect the transcript efficiency, although various mutations have different impact on the enzyme's activity.</p>
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Asunto principal:
Proteínas Recombinantes
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Análisis Mutacional de ADN
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Secuencia de Bases
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Chlorocebus aethiops
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Western Blotting
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Cromatografía Líquida de Alta Presión
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Células COS
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Fucosiltransferasas
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Genética
Límite:
Animals
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Humans
Idioma:
Zh
Revista:
Chinese Journal of Medical Genetics
Año:
2014
Tipo del documento:
Article