The sonic hedgehog protein not only plays a key role in early embryonic development, but also has essential effects on the adult nervous system, including neural stem cell proliferation, differentiation migration and ...The sonic hedgehog protein not only plays a key role in early embryonic development, but also has essential effects on the adult nervous system, including neural stem cell proliferation, differentiation migration and neuronal axon guidance. The N-terminal fragment of sonic hedgehog is the key functional element in this process. Therefore, this study aimed to clone and analyze the N-terminal fragment of the sonic hedgehog gene. Total RNA was extracted from the notochord of a Sprague-Dawley rat at embryonic day 9 and the N-terminal fragment of sonic hedgehog was amplified by nested reverse transcription-PCR. The N-terminal fragment of the sonic hedgehog gene was successfully cloned. The secondary and tertiary structures of the N-terminal fragment of the sonic hedgehog protein were predicted using Jpred and Phyre online.展开更多
Circular RNAs(circ RNAs)belong to a class of non-coding RNAs with diverse biological functions.However,little is known about their roles in case of pseudorabies virus(Pr V)infection.Here,we analyzed the expression pro...Circular RNAs(circ RNAs)belong to a class of non-coding RNAs with diverse biological functions.However,little is known about their roles in case of pseudorabies virus(Pr V)infection.Here,we analyzed the expression profile of host circ RNAs from a virulent Pr V type II strain DX(Pr V-DX)infected and an attenuated g E/TK deficient(g E-TK-Pr V)strain of Pr V infected PK-15 cells.Circ RNAs were identified by findcirc and analyzed with DESeq 2.Compared with the mock cells,449 differentially expressed(DE)circ RNAs(233 down-regulated and 216 up-regulated)from Pr V-DX infected and578 DE circ RNAs(331 down-regulated and 247 up-regulated)from g E-TK-Pr V infected PK-15 cells were identified.In addition,459 DE circ RNAs(164 down-regulated and 295 up-regulated)between the Pr V-DX and g E-TK-Pr V infected cells were identified.The expression patterns of 13 circ RNAs were validated by reverse transcription quantitative real-time PCR(RT-q PCR)and results were similar as of RNA-seq.The putative target mi RNA binding sites of DE circ RNAs were predicted by using mi Randa and ps Robot.The circ RNA-mi RNA-m RNA network was constructed and certain mi RNAs that have possible roles in antiviral immune response,such as mi R-210 and mi R-340,were predicted.GO and KEGG pathway analysis demonstrated that DE circ RNAs were enriched in the processes such as cellular metabolism,protein binding,RNA degradation and regulation of actin cytoskeleton.Collectively,these findings might provide the useful information for a better understanding of mechanisms underlying the interaction between Pr V-II and host cells.展开更多
基金sponsored by the Guangdong Provincial Natural Science Foundation,No.S2012010009592the Science and Technology Talent Foundation of Guangdong Provincial Natural Science Foundation,No.30900725+2 种基金the Joint Research Program by Southern Medical University-Shunde Guizhou Hospital,No.09000608the Science Foshan Municipal Key Project in Medical Sciences,No.201008063and the Shunde Medical Research Program,No.2011050
文摘The sonic hedgehog protein not only plays a key role in early embryonic development, but also has essential effects on the adult nervous system, including neural stem cell proliferation, differentiation migration and neuronal axon guidance. The N-terminal fragment of sonic hedgehog is the key functional element in this process. Therefore, this study aimed to clone and analyze the N-terminal fragment of the sonic hedgehog gene. Total RNA was extracted from the notochord of a Sprague-Dawley rat at embryonic day 9 and the N-terminal fragment of sonic hedgehog was amplified by nested reverse transcription-PCR. The N-terminal fragment of the sonic hedgehog gene was successfully cloned. The secondary and tertiary structures of the N-terminal fragment of the sonic hedgehog protein were predicted using Jpred and Phyre online.
基金supported by the National Key Research&Development Program of China(2016YFD0500102)Key Research&Development Program of Zhejiang Province(Grant No.2020C02011)the Fundamental Research Funds for the Central Universities(2017FZA6018)
文摘Circular RNAs(circ RNAs)belong to a class of non-coding RNAs with diverse biological functions.However,little is known about their roles in case of pseudorabies virus(Pr V)infection.Here,we analyzed the expression profile of host circ RNAs from a virulent Pr V type II strain DX(Pr V-DX)infected and an attenuated g E/TK deficient(g E-TK-Pr V)strain of Pr V infected PK-15 cells.Circ RNAs were identified by findcirc and analyzed with DESeq 2.Compared with the mock cells,449 differentially expressed(DE)circ RNAs(233 down-regulated and 216 up-regulated)from Pr V-DX infected and578 DE circ RNAs(331 down-regulated and 247 up-regulated)from g E-TK-Pr V infected PK-15 cells were identified.In addition,459 DE circ RNAs(164 down-regulated and 295 up-regulated)between the Pr V-DX and g E-TK-Pr V infected cells were identified.The expression patterns of 13 circ RNAs were validated by reverse transcription quantitative real-time PCR(RT-q PCR)and results were similar as of RNA-seq.The putative target mi RNA binding sites of DE circ RNAs were predicted by using mi Randa and ps Robot.The circ RNA-mi RNA-m RNA network was constructed and certain mi RNAs that have possible roles in antiviral immune response,such as mi R-210 and mi R-340,were predicted.GO and KEGG pathway analysis demonstrated that DE circ RNAs were enriched in the processes such as cellular metabolism,protein binding,RNA degradation and regulation of actin cytoskeleton.Collectively,these findings might provide the useful information for a better understanding of mechanisms underlying the interaction between Pr V-II and host cells.
