Lewis X(Le x) is stage specifically expressed on the surface of mammalian embryos. Histochemical evidences have showed that Le x is expressed in endometrium, and may be controlled by ovarian hormones. It has been repo...Lewis X(Le x) is stage specifically expressed on the surface of mammalian embryos. Histochemical evidences have showed that Le x is expressed in endometrium, and may be controlled by ovarian hormones. It has been reported that FUT9 gene is the real specific gene encoding α1, 3 fucosyltransferase for the synthesis of terminal fucose of Le x. In this article, the transcription of FUT9 and expression of Le x oligosaccharide antigen in mouse endometrium were investigated. The mice(22～25 g) were selected and treated as follows:(1) Estrous: females were checked by vaginal smears to determine their stage of estrus;(2) Early pregnancy: females were mated with males(1∶1), and the day of vaginal plug appearance was considered as day 1 of pregnancy;(3) Hormone replacement: after ovariectomy, mice were given subcutaneously estradiol benzoate and progesterone daily for 4 days. The results showed that the transcription level of FUT9 gene was:(1) periodically changed during estrous cycle: highest at estrus stage, and lowest at metestrus stage;(2) decreased obviously during early pregnancy, then maintained at a lower level from day 3 to day 5;(3) increased in the estradiol treatment group, and decreased obviously in the progesterone treatment group, and was at a middle level in the group treated with both estradiol and progesterone;(4) level of Le x carrying proteins on the surface of endometrium did it coincide fully with that of FUT9 gene transcription, which indicated that other factors may be also involved in the regulation of Le x expression in addition to the hormones. In summary, the expression of FUT9 gene in mouse endometrium is regulated by ovarian hormones, being up regulated by estrogen and down regulated by progesterone.展开更多
唾液酸化路易斯-X(sialyl lewis x,Slex)是选择素家族的一个共同糖配体,通过与选择素竞争性地结合炎性细胞,可以抑制炎症反应。克隆表达Slex合成过程中的关键酶,就可以在体外进行Slex的生物合成,从而进行相关生物制剂的开发。α-1,3-岩...唾液酸化路易斯-X(sialyl lewis x,Slex)是选择素家族的一个共同糖配体,通过与选择素竞争性地结合炎性细胞,可以抑制炎症反应。克隆表达Slex合成过程中的关键酶,就可以在体外进行Slex的生物合成,从而进行相关生物制剂的开发。α-1,3-岩藻糖基转移酶(alpha-(1,3)-fucosyltransferase,FT)就是参与Slex生物合成过程的关键酶之一。利用相关软件对牛的FT基因进行了生物信息学的分析,了解了FT的相关理化性质。通过PCR的方法获得了FT基因,构建了重组质粒pMD19-FT,并亚克隆至表达载体pPIC9K。通过电转化将线性化的表达质粒pPIC9K-FT整合到宿主菌Pichia pastoris GS115基因组上,构建了重组酵母GS115-FT。经诱导表达后,SDS-PAGE检测到了目的蛋白质条带,证明了此基因在P.pastoris GS115中能够可溶性表达。展开更多
文摘Lewis X(Le x) is stage specifically expressed on the surface of mammalian embryos. Histochemical evidences have showed that Le x is expressed in endometrium, and may be controlled by ovarian hormones. It has been reported that FUT9 gene is the real specific gene encoding α1, 3 fucosyltransferase for the synthesis of terminal fucose of Le x. In this article, the transcription of FUT9 and expression of Le x oligosaccharide antigen in mouse endometrium were investigated. The mice(22～25 g) were selected and treated as follows:(1) Estrous: females were checked by vaginal smears to determine their stage of estrus;(2) Early pregnancy: females were mated with males(1∶1), and the day of vaginal plug appearance was considered as day 1 of pregnancy;(3) Hormone replacement: after ovariectomy, mice were given subcutaneously estradiol benzoate and progesterone daily for 4 days. The results showed that the transcription level of FUT9 gene was:(1) periodically changed during estrous cycle: highest at estrus stage, and lowest at metestrus stage;(2) decreased obviously during early pregnancy, then maintained at a lower level from day 3 to day 5;(3) increased in the estradiol treatment group, and decreased obviously in the progesterone treatment group, and was at a middle level in the group treated with both estradiol and progesterone;(4) level of Le x carrying proteins on the surface of endometrium did it coincide fully with that of FUT9 gene transcription, which indicated that other factors may be also involved in the regulation of Le x expression in addition to the hormones. In summary, the expression of FUT9 gene in mouse endometrium is regulated by ovarian hormones, being up regulated by estrogen and down regulated by progesterone.
