Molecular cloning and expression analysis of Bmrbp1 , the Bombyx mori homologue of the Drosophila gene rbp1.
RBP1 is an important splicing factor involved in alternative splicing of the pre-mRNA of Drosophila sex-determining gene dsx. In this work, the Bombyx mori homologue of the rbp1 gene, Bmrbp1, was cloned. The pre-mRNA of Bmrbp1 gene is alternatively spliced to produce four mature mRNAs, named Bmrbp1-PA, Bmrbp1-PB, Bmrbp1-PC and Bmrbp1-PD, with nucleotide lengths of 799 nt, 1,316 nt, 894 nt and 724 nt, coding for 142 aa, 159 aa, 91 aa and 117 aa, respectively. BmRBP1-PA and BmRBP1-PD contain a N terminal RNA recognization motif (RRM) and a C terminal arginine/serine-rich domain, while BmRBP1-PB and BmRBP1-PC only share a RRM. Amino acid sequence alignments showed that BmRBP1 is conserved with its homologues in other insects and with other SR family proteins. The RT-PCR showed that Bmrbp1-PA was strongly expressed in all examined tissues and development stages, but Bmrbp1-PB was weakly expressed in these tissues and stages. The expression of both Bmrbp1-PA and Bmrbp1-PB showed no obvious sex difference. While the Bmrbp1-PC and Bmrbp1-PD were beyond detection by RT-PCR very likely due to their tissue/stage specificity. These results suggested that Bmrbp1 should be a member of SR family splicing factors, whether it is involved in the sex-specific splicing of Bmdsx pre-mRNA needs further research.
Wang
Zilong
Z
The Key Sericultural Laboratory of Agricultural Ministry, College of Sericulture and Biotechnology, Southwest University, 400716, Beibei, Chongqing, China.
Zha
Xingfu
X
He
Ningjia
N
Xiang
Zhonghuai
Z
Xa
Qingyou
Q
D017398:Alternative Splicing; D000818:Animals; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000096:biosynthesis* / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics*
D029721:Drosophila Proteins; C083909:RBP1 protein, Drosophila; D000072260:RNA Splicing Factors; D016601:RNA-Binding Proteins