中国实验动物学报
中國實驗動物學報
중국실험동물학보
ACTA LABORATORIUM ANIMALIS SCIENTIA SINICA
2014年
6期
1-8
,共8页
高建平%李玩生%曾爽%房永祥%冯海燕%杨孝朴%景志忠
高建平%李玩生%曾爽%房永祥%馮海燕%楊孝樸%景誌忠
고건평%리완생%증상%방영상%풍해연%양효박%경지충
合作小型猪%TCRα基因%生物信息学%多样性
閤作小型豬%TCRα基因%生物信息學%多樣性
합작소형저%TCRα기인%생물신식학%다양성
Hezuo minipig%Gene of TCRαchain%Bioinformatics%Diversity%Genetics
目的:探讨猪TCR基因分子结构的复杂性及其与人类的相似性。方法以公开的猪TCRα链基因为参考序列设计两对特异性引物,用RT-PCR法从合作小型猪外周血、淋巴结和脾脏的淋巴细胞中克隆了93个猪TCRα基因(简称STA)。结果测序分析表明,克隆的猪TCRα链的基因均含有可变的信号肽区和V区、高变的J区和恒定的C区的基因片段,但基因间的核苷酸序列组成都不完全相同,且具有十分复杂的多态性和多样性,基因间的同源性在68.4%~98.7%,这与TCRα链的基本基因结构特征相一致。依据TCRα基因的同源性对其分子结构、遗传演化关系和归类分析发现,在其信号肽区、FR1区和CDR1区、FR2区和CDR2区以及FR3区和CDR3区都存在一些变异集中点和变异热点区。用IMGT/V-QUEST分析方法可将合作小型猪TCRαV区( STAV)、J区( STAJ)基因片段与人类的进行比较分析,发现合作小型猪TCRα与人类的遗传演化关系较近,每个序列都能找到与人类对应的TRAV、TRAJ基因片段,甚至V区的相似性可达92%以上。结论近交培育的合作小型猪在正常状态具有应对外界复杂微生物等环境的TCR遗传多样性分子基础,且适合作为人类免疫学及疾病研究的动物模型。
目的:探討豬TCR基因分子結構的複雜性及其與人類的相似性。方法以公開的豬TCRα鏈基因為參攷序列設計兩對特異性引物,用RT-PCR法從閤作小型豬外週血、淋巴結和脾髒的淋巴細胞中剋隆瞭93箇豬TCRα基因(簡稱STA)。結果測序分析錶明,剋隆的豬TCRα鏈的基因均含有可變的信號肽區和V區、高變的J區和恆定的C區的基因片段,但基因間的覈苷痠序列組成都不完全相同,且具有十分複雜的多態性和多樣性,基因間的同源性在68.4%~98.7%,這與TCRα鏈的基本基因結構特徵相一緻。依據TCRα基因的同源性對其分子結構、遺傳縯化關繫和歸類分析髮現,在其信號肽區、FR1區和CDR1區、FR2區和CDR2區以及FR3區和CDR3區都存在一些變異集中點和變異熱點區。用IMGT/V-QUEST分析方法可將閤作小型豬TCRαV區( STAV)、J區( STAJ)基因片段與人類的進行比較分析,髮現閤作小型豬TCRα與人類的遺傳縯化關繫較近,每箇序列都能找到與人類對應的TRAV、TRAJ基因片段,甚至V區的相似性可達92%以上。結論近交培育的閤作小型豬在正常狀態具有應對外界複雜微生物等環境的TCR遺傳多樣性分子基礎,且適閤作為人類免疫學及疾病研究的動物模型。
목적:탐토저TCR기인분자결구적복잡성급기여인류적상사성。방법이공개적저TCRα련기인위삼고서렬설계량대특이성인물,용RT-PCR법종합작소형저외주혈、림파결화비장적림파세포중극륭료93개저TCRα기인(간칭STA)。결과측서분석표명,극륭적저TCRα련적기인균함유가변적신호태구화V구、고변적J구화항정적C구적기인편단,단기인간적핵감산서렬조성도불완전상동,차구유십분복잡적다태성화다양성,기인간적동원성재68.4%~98.7%,저여TCRα련적기본기인결구특정상일치。의거TCRα기인적동원성대기분자결구、유전연화관계화귀류분석발현,재기신호태구、FR1구화CDR1구、FR2구화CDR2구이급FR3구화CDR3구도존재일사변이집중점화변이열점구。용IMGT/V-QUEST분석방법가장합작소형저TCRαV구( STAV)、J구( STAJ)기인편단여인류적진행비교분석,발현합작소형저TCRα여인류적유전연화관계교근,매개서렬도능조도여인류대응적TRAV、TRAJ기인편단,심지V구적상사성가체92%이상。결론근교배육적합작소형저재정상상태구유응대외계복잡미생물등배경적TCR유전다양성분자기출,차괄합작위인류면역학급질병연구적동물모형。
Objective To analyze the complexity of molecular structure in porcine T cell receptor gene and its similarity compared to humans.Method Based on the gene of porcine T cell receptor alpha chain ( TCRα) from the Gen-Bank database, 93 swine T cell receptor alpha chain genes ( STA) were cloned by reverse transcription-polymerase chain reaction (RT-PCR) from porcine peripheral blood lymphocytes, lymph nodes and spleen.Result Sequence analysis showed that STA genes all contain a domain of variable signal peptide and V, hypervariable J and conservative C.Howev-er, nucleotide sequence of STA was not completely identical with only 68.4%to 98.7%homology among genes, and had extremely sophisticated polymorphism and diversity.This was accord with the genetic structure of TCRαchain.Molecular structure, genetic evolution and classification of these genes were carried out according to the homology of TCRαgene, which all have several sites and zones of mutation on the domain of signal peptide, FR1 and CDR1, FR2 and CDR2, FR3 and CDR3.Analysis of similarity and classification of TCRαV domain(STAV)and J domain (STAJ) of Hezuo minipig u-sing IMGT/V-QUEST tools compared with those of humans found the genetic evolution relationship that was closer, and each of TRAV and TRAJ also found to have a corresponding fragment of humans, ever in 92% of similarity of TRAV be-tween swine and humans.Conclusion Our results indicate that inbred Hezuo minipig possesses genetic diversity against complicated environment of microbes in healthy status, and Hezuo minipig is suitable as an animal model for research on human immunology and diseases.