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利用聚合酶链反应-抑制消减杂交技术鉴定无瘢痕伤口愈合中差异表达的基因。

Identification of differentially expressed genes in scarless wound healing utilizing polymerase chain reaction-suppression subtractive hybridization.

作者信息

Kathju Sandeep, Satish Latha, Rabik Cara, Rupert Terra, Oswald Duane, Johnson Sandra, Hu Fen Ze, Post J Christopher, Ehrlich Garth D

机构信息

Center for Genomic Sciences, Allegheny Singer Research Institute, Allegheny General Hospital, Pittsburgh, Pennsylvania 15212-4772, USA.

出版信息

Wound Repair Regen. 2006 Jul-Aug;14(4):413-20. doi: 10.1111/j.1743-6109.2006.00140.x.

Abstract

Wound healing in fetal skin is well known to proceed without scarring, whereas adult (postnatal) skin wound healing is accompanied by scar formation. To identify differentially expressed genes during fetal wound (FW) healing, we have used polymerase chain reaction-suppression subtractive hybridization. This technique allows for a comparative analysis across the entire transcriptome of FW vs. unwounded fetal control tissue, including even potentially novel sequences. Our subtractive hybridization protocol identified 15 clones that are overexpressed in healing FWs, and 20 clones that are underexpressed. These include genes with both known and unknown functions. We have confirmed the differential pattern of expression for four of these candidate genes: elongation factor 1 alpha, elongation initiation factor 4e, and two transcripts thus far known only as an expressed sequence tags. With this approach, we have also identified novel genes potentially involved in scarless wound healing.

摘要

众所周知,胎儿皮肤伤口愈合过程中不会形成瘢痕,而成人(出生后)皮肤伤口愈合则伴有瘢痕形成。为了鉴定胎儿伤口(FW)愈合过程中差异表达的基因,我们采用了聚合酶链反应抑制消减杂交技术。该技术能够对FW与未受伤胎儿对照组织的整个转录组进行比较分析,甚至包括潜在的新序列。我们的消减杂交方案鉴定出15个在愈合的FW中过度表达的克隆,以及20个表达不足的克隆。这些克隆包括功能已知和未知的基因。我们已经证实了其中四个候选基因的差异表达模式:延伸因子1α、延伸起始因子4e,以及两个迄今仅作为表达序列标签已知的转录本。通过这种方法,我们还鉴定出了可能参与无瘢痕伤口愈合的新基因。

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