Expression of tyrosinase-like protein genes and their functional analysis in melanin synthesis of Pacific Oyster (Crassostrea gigas).
Expression of tyrosinase-like protein genes and their functional analysis in melanin synthesis of Pacific Oyster (Crassostrea gigas).
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DOI：
10.1016/j.gene.2022.146742
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发表时间：
2022-07
期刊：
影响因子：
3.5
通讯作者：
Yijing Zhu;Qi Li;Hong Yu;Shikai Liu;L. Kong
中科院分区：
文献类型：
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作者：
Yijing Zhu;Qi Li;Hong Yu;Shikai Liu;L. Kong
软体动物的颜色多态性对消费者的偏好非常感兴趣。尽管壳颜色变异的遗传性是通过实验杂交进行的，但人们对这些模式所涉及的分子基础知之甚少。酪氨酸酶样蛋白是重要的酶，是3型铜蛋白超家族的成员。本研究在太平洋牡蛎巨牡蛎中鉴定出两个酪氨酸酶样蛋白基因，包括CgTyp-1和CgTyp-3。组织表达分析表明CgTyp-1和CgTyp-3主要在地幔中表达。特别是，无论是在左侧还是右侧地幔，它们在边缘地幔中的表达均显着高于在中央地幔中的表达。另外，CgTyp-1和CgTyp-3主要在黑壳色牡蛎中表达，在白壳色牡蛎中表达量相对较低。原位杂交表明，无论是黑壳色牡蛎还是白壳色牡蛎，在外皱襞的外上皮内均检测到CgTyp-1和CgTyp-3的阳性信号。干扰后，CgTyp-1和CgTyp-3mRNA的表达水平显着减弱，RNAi效率分别达到84.72%和71.58%。此外，敲低CgTyp-1或CgTyp-3，明显降低了外套膜的酪氨酸酶活性。此外，通过沉默eachTyp，外皱襞上皮内黑素体的数量急剧减少。这些发现表明CgTyp-1和CgTyp-3可能参与黑色素合成，这有助于深入了解CgTyp-1和CgTyp-3壳色素沉着的调节机制。千兆。
Color polymorphism in Mollusca is of great interest for consumer preference. Although the heritability of shell color variation has been conducted by experimental crossing, little is known about molecular basis involved in these patterns. Tyrosinase-like proteins are important enzymes which are members of the type-3 copper protein superfamily. In this research, two tyrosinase-like protein genes includingCgTyp-1andCgTyp-3were identified in the Pacific oysterCrassostrea gigas. Tissue expression analysis showed thatCgTyp-1andCgTyp-3were dominantly expressed in the mantle. Particularly, they were expressed significantly higher in the edge mantle than that in the central mantle whether on the left or right mantles. Additionally, expressions ofCgTyp-1andCgTyp-3were mainly found in the black shell color oysters, with relative lower levels in the white shell color oysters. In situ hybridization showed that positive signals forCgTyp-1andCgTyp-3were both detected within the outer epithelium of the outer fold either in the black or white shell color oysters. After interference, the expression levels ofCgTyp-1andCgTyp-3mRNA were significantly attenuated, and the efficiency of RNAi reached 84.72% and 71.58%, respectively. Besides, knockdownCgTyp-1orCgTyp-3, obviously decreased the tyrosinase activity of mantles. Furthermore, the number of the melanosomes within epithelium of the outer fold was sharply reduced by silencing of eachTyp. These findings argue thatCgTyp-1andCgTyp-3may be involved in the melanin synthesis, which lends insight into regulation mechanism of shell pigmentation inC. gigas.