中文摘要：

collagenⅥ（COLⅥ）是胶原蛋白家族的重要成员,广泛分布于动物体细胞外基质,在脊椎动物骨骼形成过程中发挥重要作用。为了探究COLⅥ在马氏珠母贝贝壳形成中的作用,本研究运用聚合酶链式反应（PCR）和c DNA末端快速扩增技术（RACE）获得了马氏珠母贝Pm COLVIA6基因的全长序列,克隆获得其启动子区域,并对其蛋白结构进行了分析。荧光定量PCR技术检测了该基因在马氏珠母贝各个组织的表达量分布,并利用原位杂交技术检测Pm COLVIA6基因在外套膜的表达定位。结果表明：Pm COLVIA6序列全长为2 100 bp,开放阅读框（ORF）长为1 875 bp,编码624个氨基酸,5’非翻译区（5’UTR）长56 bp,3’非翻译区（3’UTR）长169 bp,于5’调控区克隆得到长为1 826 bp的序列,可能存在4个启动子序列。Pm COLVIA6蛋白含有典型的v WA结构域,但三股螺旋结构缺失。荧光定量分析表明Pm COLVIA6基因在马氏珠母贝的各个组织均有表达,而在珍珠囊和外套膜中央区的表达量显著高于其它组织。原位杂交结果显示Pm COLVIA6的m RNA主要分布于外套膜中央区的外表皮细胞。综上所述,Pm COLVIA6可能参与马氏珠母贝珍珠层的形成。

英文摘要：

Type Ⅵ collagen（COLⅥ） is a vital member of the collagen family and it is ubiquitously expressed throughout all ECMs in animals, which plays important roles in bone formation of vertebrates. In this study, to investigate the function of COLⅥ in shell formation of Pinctada martensii, using polymerase chain reaction and rapid amplification of c DNA ends technology, the full length of Pm COLVIA6 gene was obtained from Pinctada martensii, then we amplificated the promoter region of Pm COLVIA6 and analysised its protein structure. Real-time PCR was used to detect the tissue-specific expression of this gene in Pinctada martensii, and situ hybridization was used to detect the express location of it. The results showed the total length of Pm COLVIA6 gene was 2 100 bp,including 1 875 bp of the open reading frame（ORF） which encoding 624 amino acids, a 5’ UTR of 56 bp and a 3’UTR of 169 bp. The promoter region was cloned with 1 826 bp region in the upstream of transcription initiation site, which might include four predicted promoter sequences. Pm COLVIA6 obtained the conserved v WA domain but lacked the triple-helical structure. Expression pattern of Pm COLVIA6 in different tissues was tested by Real-Time PCR. Pm COLVIA6 was constitutively expressed in all studied tissues in Pinctada martensii with the significantly high expression in pearl sac and in mantle center. In situ hybridization experiment, we found that m RNA of Pm COLVIA6 gene specifically expressed in the outer of mantle center epithelium. These studies indicated that Pm COLVIA6 might regulate nacre formation of Pinctada martensii.