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ＦＤ耐热逆转录酶（ＦＤ－ＴＲＴ）来自一株栖热杆菌菌株．通过ＲＴ－ＰＣＲ方法，论文对ＦＤ－ＴＲＴ的部分酶学性质进行了研究．ＦＤ耐热逆转录酶能耐受９５℃的高温，最适反应温度在６５～７５℃左右，这时ＲＮＡ的高级结构能解开，可以提高逆转录反应的效率；同时引物和ＲＮＡ模板识别的专一性强，可大大提高逆转录的特异性．实验表明ＦＤ－ＴＲＴ逆转录反应的最适条件为：２５ｍｍｏｌ／ＬＴｒｉｓ－ＨＣｌ（ｐＨ８．８，１５ｍｍｏｌ／Ｌ（ＮＨ４）２ＳＯ４，１００μｇ／ｍｌ明胶，５００μｍｏｌ／ＬｄＮＴＰｓ，２５ｐｍｏｌ逆转录引物，１ｍｍｏｌ／ＬＭｎＣｌ２，２ＵＦＤ耐热逆转录酶，反应在６５～７０℃下进行．在上述条件下，用ＲＴ－ＰＣＲ可检出少于５ｐｇ外周血总ＲＮＡ中特异的α珠蛋白ｍＲＮＡ．
FD heat-resistant reverse transcriptase (FD-TRT) is from a strain of Thermus. The partial enzymatic properties of FD-TRT were studied by RT-PCR. FD heat-resistant reverse transcriptase can withstand high temperatures of 95 ℃, the optimal reaction temperature is about 65 ~ 75 ℃, then RNA’s high-level structure can be solved, can improve the efficiency of reverse transcription reaction; the same time, primers and RNA template recognition Specificity, can greatly enhance the specificity of reverse transcription. The optimal conditions for the reverse transcription reaction of FD-TRT were 25mmol / L Tris-HCl (pH8.8, 15mmol / L (NH4) 2SO4, 100μg / ml gelatin, 500μmol / L dNTPs, 25pmol reverse transcription primer, 2UFD heat-resistant reverse transcriptase and the reaction was carried out at 65-70 ° C. Under these conditions, less than 5 μg of specific α-globin mRNA in peripheral blood total RNA was detected by RT-PCR.