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Gene-clone and expression of Barx1 in different tooth of the mini-pig at embryonic day 40

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Author:
No author available
Journal Title:
Chinese Journal of Stomatology
Issue:
10
DOI:
10.3760/cma.j.issn.1002-0098.2012.10.011
Key Word:
猪,雏型;胚胎和胎儿发育;显微切割;Barx1;Swine,miniature; Embryonic and fetal development; Microdissection; Barx1

Abstract: Objective To partially clone and compare the quantitative expression of tooth development-related gene Barx1 in different teeth of the mini-pig embryo at embryonic day 40,and to investigate the relationship between Barxl spatial quantitative expression and tooth morphogenesis.Methods The mini-pig Barx1 genes was partially cloned and the mRNA sequences of human Barx1 genes was aligned with expressed sequence tags (EST) of pig by basic local alignment search tool (BLAST),which were assembled with DNAman v5.2.2. With designed primers,Barxl was partially cloned in use of reverse transcription polymerase chain reaction (PCR),and tested by BLAST with all the species in NCBI database and confirmed as one part of target gene. Laser capture microdissection was used to collect tooth samples from frozen sections which were prepared before in -80 ℃ freezer. Real-time PCR was carried out to analyze quantitative expression in different teeth. Results Partial mini-pig Barx1 gene of 698 bp was cloned.Real-time PCR showed that,glyceraldehyde-3-phosphate dehydrogenase used as loading control,the figures of 2 -△CT of lower deciduous incisor,canine,the third premolar and molar were 0.000 249,0.000 715,0.026 096 and 0.112 656,respectively.There was a trend of increasing expression from anterior to posterior teeth.Conclusions Barx1 gene could be related to the number or differentiation of tooth cusps.

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