NDLI: Rapid cloning and bioinformatic analysis of spinach Y chromosome-specific EST sequences

Content Provider	Springer Nature Link
Author	DENG, CHUAN LIANG ZHANG, WEI LI CAO, YING WANG, SHAO JING LI, SHU FEN GAO, WU JUN LU, LONG DOU
Copyright Year	2015
Abstract	The genome of spinach single chromosome complement is about 1000 Mbp, which is the model material to study the molecular mechanisms of plant sex differentiation. The cytological study showed that the biggest spinach chromosome (chromosome 1) was taken as spinach sex chromosome. It had three alleles of sex-related X, X $^{ m }$ and Y. Many researchers have been trying to clone the sex-determining genes and investigated the molecular mechanism of spinach sex differentiation. However, there are no successful cloned reports about these genes. A new technology combining chromosome microdissection with hybridization-specific amplification (HSA) was adopted. The spinach Y chromosome degenerate oligonucleotide primed-PCR (DOP-PCR) products were hybridized with cDNA of the male spinach flowers in florescence. The female spinach genome was taken as blocker and cDNA library specifically expressed in Y chromosome was constructed. Moreover, expressed sequence tag (EST) sequences in cDNA library were cloned, sequenced and bioinformatics was analysed. There were 63 valid EST sequences obtained in this study. The fragment size was between 53 and 486 bp. BLASTn homologous alignment indicated that 12 EST sequences had homologous sequences of nucleic acids, the rest were new sequences. BLASTx homologous alignment indicated that 16 EST sequences had homologous protein-encoding nucleic acid sequence. The spinach Y chromosome-specific EST sequences laid the foundation for cloning the functional genes, specifically expressed in spinach Y chromosome. Meanwhile, the establishment of the technology system in the research provided a reference for rapid cloning of other biological sex chromosome-specific EST sequences.
Starting Page	705
Ending Page	713
Page Count	9
File Format	PDF
ISSN	00221333
Journal	Journal of Genetics
Volume Number	94
Issue Number	4
e-ISSN	09737731
Language	English
Publisher	Springer India
Publisher Date	2015-12-14
Publisher Institution	Indian Academy of Sciences
Publisher Place	New Delhi
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	spinach Y chromosome microdissection hybridization-specific amplification expressed sequence tag Life Sciences Microbial Genetics and Genomics Plant Genetics & Genomics Animal Genetics and Genomics Evolutionary Biology
Content Type	Text
Resource Type	Article
Subject	Genetics

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