摘要
层粘连蛋白是重要的细胞外基质成分之一,在基膜的构建及细胞的黏附等方面都有重要作用。通过检索微孢子虫数据库发现家蚕微孢子虫(Nosema bombycis)存在假定层粘连蛋白基因Nb HLAMB4,根据该基因序列设计引物,以家蚕微孢子虫基因组DNA为模板克隆了Nb HLAMB4基因。该基因的开放阅读框(ORF)全长为1 104 bp,预测编码的蛋白质含有367个氨基酸,分子质量为43.74 k D,等电点为5.72。Nb HLAMB4蛋白序列含有一个N-糖基化位点和Pfam:SAS-6_N结构域,无信号肽,也无跨膜结构域。应用酵母双杂交技术,以Nb HLAMB4作为诱饵蛋白,通过筛选家蚕中肠c DNA文库,初步得到3个与Nb HLAMB4互作的候选蛋白质,分别是家蚕热休克蛋白(Hsp40)、锌指蛋白420类似物(similar to zinc finger protein 420)、氨肽酶N(aminopeptidase N)。研究结果为阐明家蚕微孢子虫的侵染机制提供了新的线索。
Laminin is one of the important components of the extracellular matrix with crucial roles in the construction of basement membrane and cell adhesion etc. The hypothetical laminin gene Nb HLAMB4 was found in Nosema bombycis by searching the microsporidian sequence database. Primer sequences were designed to clone Nb HLAMB4 from the genomic DNA of N. bombycis according to its gene sequence. Sequence analysis showed that Nb HLAMB4 gene contained a complete ORF of 1 104 bp encoding a polypeptide of 367 amino acids with a molecular weight of 43. 74 kD and an isoelectric point of 5. 72. A N-glycosylation site and Pfam：SAS-6_N domain were predicted in Nb HLAMB4 protein sequence,but it has no signal peptide or transmembrane domain. Three candidate proteins interacting with Nb HLAMB4,namely Hsp40 protein,protein similar to zinc finger protein 420,and aminopeptidase N,were filtered out through yeast two-hybrid system by screening of silkworm midgut c DNA library and using Nb HLAMB4 as the bait protein. These results provide new clues to clarify the infection mechanism of N.bombycis.
出处
《蚕业科学》
CAS
CSCD
北大核心
2016年第4期656-661,共6页
ACTA SERICOLOGICA SINICA
基金
现代农业产业技术体系建设专项(No.CARS-22)
