title: Identification and Characterization of 30 K Protein Genes Found in Bombyx mori (Lepidoptera: Bombycidae) Transcriptome.
abstract: The 30 K proteins, the major group of hemolymph proteins in the silkworm, Bombyx mori (Lepidoptera: Bombycidae), are structurally related with molecular masses of ∼30 kDa and are involved in various physiological processes, e.g., energy storage, embryonic development, and immune responses. For this report, known 30 K protein gene sequences were used as Blastn queries against sequences in the B. mori transcriptome (SilkTransDB). Twenty-nine cDNAs (Bm30K-1-29) were retrieved, including four being previously unidentified in the Lipoprotein_11 family. The genomic structures of the 29 genes were analyzed and they were mapped to their corresponding chromosomes. Furthermore, phylogenetic analysis revealed that the 29 genes encode three types of 30 K proteins. The members increased in each type is mainly a result of gene duplication with the appearance of each type preceding the differentiation of each species included in the tree. Real-Time Quantitative Polymerase Chain Reaction (Q-PCR) confirmed that the genes could be expressed, and that the three types have different temporal expression patterns. Proteins from the hemolymph was separated by SDS-PAGE, and those with molecular mass of ∼30 kDa were isolated and identified by mass spectrometry sequencing in combination with searches of various databases containing B. mori 30K protein sequences. Of the 34 proteins identified, 13 are members of the 30 K protein family, with one that had not been found in the SilkTransDB, although it had been found in the B. mori genome. Taken together, our results indicate that the 30 K protein family contains many members with various functions. Other methods will be required to find more members of the family.
authors.affiliation: The Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, 12 Zhongguancun South Street Beijing, 100081, China The College of Biological Sciences, China Agricultural University, Beijing, 100094, China.
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Shi
authors.affiliation: The Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, 12 Zhongguancun South Street Beijing, 100081, China.
authors.forename: Yi-Nü
authors.initials: YN
authors.lastname: Li
authors.affiliation: The Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu Province, 212018, China.
authors.forename: Yong-Zhu
authors.initials: YZ
authors.lastname: Yi
authors.affiliation: The College of Biological Sciences, China Agricultural University, Beijing, 100094, China.
authors.forename: Xing-Guo
authors.initials: XG
authors.lastname: Xiao
authors.affiliation: The Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, 12 Zhongguancun South Street Beijing, 100081, China zhifangzhang@yahoo.com.
authors.forename: Zhi-Fang
authors.initials: ZF
authors.lastname: Zhang
mesh_terms: D000818:Animals; D012831:Bombyx / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D018076:DNA, Complementary / Q000302:isolation & purification; D005786:Gene Expression Regulation; D017344:Genes, Insect; D006458:Hemolymph / Q000378:metabolism; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D020543:Proteome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D008074:Lipoproteins; D020543:Proteome
keywords: gene family; phylogenetic analysis; temporal expression pattern