Objective: To establish a sensitive, nonradioactive in situ hybridization method to detect the expression of gut regulatory peptide genes. Methods: The digoxigenin (Dig) labeled somatostatin (SS) RNA probes were synth...Objective: To establish a sensitive, nonradioactive in situ hybridization method to detect the expression of gut regulatory peptide genes. Methods: The digoxigenin (Dig) labeled somatostatin (SS) RNA probes were synthesized with in vitro transcription system and the in situ hybridization was conducted on the cryostat sections of rat stomach , duodenum and pancreas. Results: Positive reaction sites were purple-blue in color and located in the cytoplasm with a light background and a strong contrast. Both the morphosis and distribution of the SS mRNA posrtive cells corresponded well with those of the previously reported SS immunoreactive cells. Conclusion: The Dig -labeled cRNA probe in situ hybridization method is simple, sensitive and reliable for the study of gut regulatory peptide gene expression.展开更多
为了研究褪黑素受体在人脑中的表达状况以及褪黑素在老年性痴呆中的作用 ,本研究克隆了 Mel1 a和 RORβ等两种褪黑素受体的 c DNA并将其转入 p Bluescript ( KS+)质粒载体中。重组质粒通过限制性内切酶鉴定分析并测序。结果表明 ,克隆...为了研究褪黑素受体在人脑中的表达状况以及褪黑素在老年性痴呆中的作用 ,本研究克隆了 Mel1 a和 RORβ等两种褪黑素受体的 c DNA并将其转入 p Bluescript ( KS+)质粒载体中。重组质粒通过限制性内切酶鉴定分析并测序。结果表明 ,克隆的序列与 Gen Bank中的序列一致。利用线性化的质粒模板反转出 c RNA探针 。展开更多
用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经...用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经病理证实的肺癌及癌周正常肺组织新鲜标本 ,用液氮速冻 ,4%多聚甲醛固定。在冰冻切片上以 c RNA探针原位杂交进行 Fas L m RNA表达分布的观察。结果 :在鳞癌、腺癌和小细胞肺癌的癌组织及癌旁肺组织中均可检出 Fas L的杂交信号。镜下见鳞癌组织中 Fas展开更多
文摘Objective: To establish a sensitive, nonradioactive in situ hybridization method to detect the expression of gut regulatory peptide genes. Methods: The digoxigenin (Dig) labeled somatostatin (SS) RNA probes were synthesized with in vitro transcription system and the in situ hybridization was conducted on the cryostat sections of rat stomach , duodenum and pancreas. Results: Positive reaction sites were purple-blue in color and located in the cytoplasm with a light background and a strong contrast. Both the morphosis and distribution of the SS mRNA posrtive cells corresponded well with those of the previously reported SS immunoreactive cells. Conclusion: The Dig -labeled cRNA probe in situ hybridization method is simple, sensitive and reliable for the study of gut regulatory peptide gene expression.
文摘为了研究褪黑素受体在人脑中的表达状况以及褪黑素在老年性痴呆中的作用 ,本研究克隆了 Mel1 a和 RORβ等两种褪黑素受体的 c DNA并将其转入 p Bluescript ( KS+)质粒载体中。重组质粒通过限制性内切酶鉴定分析并测序。结果表明 ,克隆的序列与 Gen Bank中的序列一致。利用线性化的质粒模板反转出 c RNA探针 。
文摘用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经病理证实的肺癌及癌周正常肺组织新鲜标本 ,用液氮速冻 ,4%多聚甲醛固定。在冰冻切片上以 c RNA探针原位杂交进行 Fas L m RNA表达分布的观察。结果 :在鳞癌、腺癌和小细胞肺癌的癌组织及癌旁肺组织中均可检出 Fas L的杂交信号。镜下见鳞癌组织中 Fas
