Lynn S, Morgan J M, Lamb H K, Meissner G, Gillespie J I
Department of Physiological Sciences and Biochemistry, Medical School, University of Newcastle upon Tyne, England.
FEBS Lett. 1995 Sep 18;372(1):6-12. doi: 10.1016/0014-5793(95)00924-x.
Partial cDNAs of the ryanodine receptor were cloned using PCR analysis from reverse transcribed total and mRNA, extracted from freshly isolated pregnant, non-pregnant, and cultured human myometrial smooth muscle. The identity of these clones was confirmed by nucleotide sequencing of the fragments and indicate the expression of both the skeletal and brain ryanodine receptor isoforms in these preparations. In freshly isolated non-pregnant myometrial tissue, membrane fractions displaying specific [3H]ryanodine binding activities were isolated using density gradient centrifugation. SDS-PAGE of the sucrose gradient fractions indicated the specific comigration of a polypeptide with a molecular mass of approximately 544 kDa with the ryanodine binding activity.
利用聚合酶链反应（PCR）分析法，从新鲜分离的妊娠、未妊娠及培养的人子宫肌层平滑肌中提取的总RNA和mRNA反转录产物中克隆出兰尼碱受体的部分cDNA。通过对片段进行核苷酸测序，证实了这些克隆的身份，并表明在这些标本中同时表达了骨骼肌型和脑型兰尼碱受体亚型。在新鲜分离的未妊娠子宫肌层组织中，使用密度梯度离心法分离出显示特异性[3H]兰尼碱结合活性的膜组分。蔗糖梯度组分的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）表明，一条分子量约为544 kDa的多肽与兰尼碱结合活性特异性共迁移。
