家蚕30K蛋白BmLP1的表达模式及其在胚胎中的作用  

Expression Pattern of Silkworm 30K Protein BmLP1 and Its Role in Embryonic Development

作　　者：张艳[1] 董照明[1] 夏建华 令狐青青 万娜 张仁义 卢忠燕 林英[1] 赵萍[1] ZHANG Yan;DONG Zhaoming;XIA Jianhua;LINGHU Qingqing;WAN Na;ZHANG Renyi;LU Zhongyan;LIN Ying;ZHAO Ping(Integrative Science Center of Germplasm Creation in Western China(Chongqing)Science City,Southwest University/Biological Science Research Center,Academy for Advanced Interdisciplinary,Chongqing 400715,China)

机构地区：[1]西南大学西部(重庆)科学城种质创制大科学中心/前沿交叉学科研究院生物学研究中心,重庆400715

出　　处：《西南大学学报（自然科学版）》2024年第10期99-106,共8页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(32202750);重庆市基础与前沿研究计划项目(cstc2021jcyj-msxmX0582);重庆市创新群体项目(cstc2020jcyj-cxtt)。

摘　　要：家蚕30K蛋白是家蚕血液和卵中的高丰度蛋白,参与营养、免疫、抑制细胞凋亡等功能.探究30K蛋白在家蚕体内的表达模式有助于更清楚地了解30K蛋白的功能.利用生物信息学方法发现4种30K蛋白基因Bmlp1,Bmlp2,Bmlp3和Bmlp7都含有信号肽.其中,Bmlp1与其他30K蛋白的序列相似性小于50%.半定量RT-PCR结果表明,Bmlp1从5龄第3 d到化蛹第1 d在脂肪体中表达量都比较高,之后表达量降低.Western blotting结果表明,BmLP1从5龄第5 d到化蛹第4 d在血淋巴中的含量都比较高,之后含量降低.与血淋巴中的变化趋势不同,卵巢中的BmLP1在上蔟第2 d被检测到,之后含量一直增加,化蛾时含量达到最高.这是由于上蔟第2 d卵巢管迅速长大,血液中的BmLP1等营养蛋白逐渐转运至卵巢管的未受精卵并开始积累.免疫组织化学定位结果表明,在胚胎发育前期,BmLP1分布于蚕卵的卵黄颗粒中,在孵化前期,BmLP1被吞入胚胎的肠道内.体外孵育结果表明,蚁蚕粗提物可以降解BmLP1,暗示蚁蚕肠道内可能存在某种蛋白酶可以将大部分BmLP1水解,最终为蚁蚕的孵化提供能量或者氨基酸.系统分析了BmLP1在不同发育时期脂肪体、血液、卵巢和胚胎中的表达特征,揭示了其在胚胎发育过程中的重要作用.30K proteins are abundant in silkworm hemolymph and egg.They are involved in nutrition,immunity,inhibition of apoptosis and other functions.We studied the expression of 30K proteins to better understand their functions.In this research,bioinformatic analysis showed that all the Bmlp1,Bmlp2,Bmlp3 and Bmlp7 genes have signal peptides.The sequence similarity between Bmlp1 and other 30K protein genes is less than 50%.The result of semi-quantitative RT-PCR showed that Bmlp1 was highly expressed from day 3 of fifth instar larvae to day 1 of pupae,and then decreased.By Western blotting assay,we found that the BmLP1 antibody could only bind with BmLP1 protein,not BmLP2,BmLP3 and BmLP7 proteins,suggesting that the antibody has good specificity.The Western blotting result showed that BmLP1 was abundant from day 5 of fifth instar larvae to day 4 of pupae,and then decreased,whereas BmLP1 was firstly observed after day 2 of wandering,then increased gradually,and reached a maximum level in the moth.This is because that ovary tubes grew quickly from day 2 of wandering,and BmLP1 and other nutrients-storage proteins were transported into the ovary tubes and accumulated in the unfertilized eggs.Immmunohistochemistry result showed that BmLP1 was located in the yolk granules on day 6 of egg and in the gut before the hatching.We incubated BmLP1 with the new hatched larvae extract in vitro,and found that the extract could degraded the majority of BmLP1,indicating that there may be one protease in the gut responsible for the degradation of BmLP1.BmLP1 could provide the energy and amino acid resources for the hatching of baby silkworm larvae.This study explored the changing pattern of silkworm 30K protein BmLP1 in the fat body,hemolymph,ovary and egg at different developmental stage,laying the foundation for understanding the function of BmLP1 in the silkworm embryonic development.

关 键 词：家蚕 30K蛋白 BmLP1 胚胎发育 血淋巴 

分 类 号：S881.2[农业科学—特种经济动物饲养]