Electronic Cloning And Bioinformatics Analysis Of Genes Associated With Sugar Beet Marker Locus BvRE016

Sugar beet(Beta Vulgaris L.)is the main sugar crop in northern China.In China,sugar beet biotechnology research is relatively delayed because of a lack of gene resources,and the isolated genes are limited in few types.Obtaining more gene resources and their annotation information has become a necessary prerequisite for sugar beet biotechnology research.Through the previous research,our group had obtained an SSR molecular marker Bv RE016(patent pending),related to an important biennial trait of sugar beet.In this study,a sugar beet germplasm with positive genotyping results of this marker was used as plant material,and the complete coding region of locus-related genes was obtained by in silico isolation by virtue of the transcript sequence library collected previously by our group.Firstly,the marker site was extended in silico by combining the local transcript sequence library and the public sequence library;after the extension result was located in the genome,biological experiments were carried out to isolate the target gene fragment from sugar beet genome,and the in silico extension result was meanwhile verified,which means the in silico isolation had been accomplished.Finally,the structure and key sequences of the target gene locus in the genome are determined by bioinformatics analysis,and the structural and functional characteristics of the coding products are predicted.Through this study,annotations for the marker locus Bv RE016 and the corresponding biennial trait of sugar beet would be provided,and more reliable sugar beet new sequence information would be obtained,which will provide sequence materials and gene materials for sugar beet biotechnology theory research.Combined with the follow-up functional analysis,it will lay a foundation for sugar beet gene resource mining and other biotechnology research.Main research results:(1)Through genome mapping and in silico extension,the marker site Bv RE016 was determined to be located inside a coding region on chromosome 9,and the coding product belongs to the neuropeptide-like protein(NLP)family.(2)By PCR verification,a band around 5000 bp was obtained,which was consistent with the result of in silico extension.The target fragment was isolated from the genome and obtained a sequence of 4386 bp,including a complete coding region,and the gene was named as Bv NLP2-like.(3)Using bioinformatics tools,it was predicted that the gene coding product Bv NLP2-like has the hydrophilic physical property,one transmembrane domain and the characteristic domain of NLP family.(4)Three exons were predicted to be in the gene locus in the genome,and the splice site verification scheme as well as the required primers were designed.In addition,a variety of cis-regulatory elements were excavated upstream.