中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
42期
6769-6774
,共6页
霍丽蓉%王兰英%邹俊华%钟南
霍麗蓉%王蘭英%鄒俊華%鐘南
곽려용%왕란영%추준화%종남
组织构建%组织工程%核不均一核糖核蛋白E1%表达载体%神经细胞%国家自然科学基金
組織構建%組織工程%覈不均一覈糖覈蛋白E1%錶達載體%神經細胞%國傢自然科學基金
조직구건%조직공정%핵불균일핵당핵단백E1%표체재체%신경세포%국가자연과학기금
ribonucleoprotein,heterogeneous%neurons%transfection
背景:人源核不均一核糖核蛋白E1功能广泛,可参与神经系统骨架蛋白的表达。目的:为深入研究其在神经细胞中的作用,构建其真核表达载体,观察其在神经细胞中的表达。方法:利用真核表达载体pcDNATM4/His C,通过亚克隆构建核不均一核糖核蛋白E1-pcDNATM 4/His C重组质粒,经酶切、测序鉴定,通过转染神经细胞SH-SY5Y,采用western-blot,RT-PCR鉴定核不均一核糖核蛋白E1重组质粒的表达,并观察转染细胞的生长现象。结果与结论:成功构建了核不均一核糖核蛋白 E1的真核表达载体,mRNA 和蛋白水平上均证实了该质粒可在神经细胞SH-SY5Y中正确表达。SH-SY5Y细胞在转染核不均一核糖核蛋白E1后表现为加速生长。提示该蛋白对神经细胞的生长发育具有重要的作用。该载体为进一步研究核不均一核糖核蛋白E1在神经系统中的功能提供了前提条件。
揹景:人源覈不均一覈糖覈蛋白E1功能廣汎,可參與神經繫統骨架蛋白的錶達。目的:為深入研究其在神經細胞中的作用,構建其真覈錶達載體,觀察其在神經細胞中的錶達。方法:利用真覈錶達載體pcDNATM4/His C,通過亞剋隆構建覈不均一覈糖覈蛋白E1-pcDNATM 4/His C重組質粒,經酶切、測序鑒定,通過轉染神經細胞SH-SY5Y,採用western-blot,RT-PCR鑒定覈不均一覈糖覈蛋白E1重組質粒的錶達,併觀察轉染細胞的生長現象。結果與結論:成功構建瞭覈不均一覈糖覈蛋白 E1的真覈錶達載體,mRNA 和蛋白水平上均證實瞭該質粒可在神經細胞SH-SY5Y中正確錶達。SH-SY5Y細胞在轉染覈不均一覈糖覈蛋白E1後錶現為加速生長。提示該蛋白對神經細胞的生長髮育具有重要的作用。該載體為進一步研究覈不均一覈糖覈蛋白E1在神經繫統中的功能提供瞭前提條件。
배경:인원핵불균일핵당핵단백E1공능엄범,가삼여신경계통골가단백적표체。목적:위심입연구기재신경세포중적작용,구건기진핵표체재체,관찰기재신경세포중적표체。방법:이용진핵표체재체pcDNATM4/His C,통과아극륭구건핵불균일핵당핵단백E1-pcDNATM 4/His C중조질립,경매절、측서감정,통과전염신경세포SH-SY5Y,채용western-blot,RT-PCR감정핵불균일핵당핵단백E1중조질립적표체,병관찰전염세포적생장현상。결과여결론:성공구건료핵불균일핵당핵단백 E1적진핵표체재체,mRNA 화단백수평상균증실료해질립가재신경세포SH-SY5Y중정학표체。SH-SY5Y세포재전염핵불균일핵당핵단백E1후표현위가속생장。제시해단백대신경세포적생장발육구유중요적작용。해재체위진일보연구핵불균일핵당핵단백E1재신경계통중적공능제공료전제조건。
BACKGROUND:The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE:To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS:Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION:We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.