摘要
利用ＲＴ－ＰＣＲ技术分离低密度脂蛋白受体基因ｃＤＮＡ，将长为２６０５ｂｐ的ｃＤＮＡ插入质粒ｐＪＮ６，并经全序列测定证实，该序列与已报道序列相比，存在两个变异碱基，即第７５４位Ｃ→Ｔ，和１６５４位的Ｇ→Ａ，这两个变异碱基并不改变编码的氨基酸，利用ＬＤＬ受体基因ｃＤＮＡ中的ＣｌａⅠ片段作为探针，检测到ＬＤＬ受体基因上外显子８的ＳｔｕⅠ位点是个限制性片段长度多态性位点。所克隆的ｃＤＮＡ含有可译框架的全部密码，因此可作为基因表达材料。
A cDNA containing the entire coding region for LDL receptor was obtained by reverse transcription and polymerase chain reaction (RT-PCR). The cDNA was inserted into vector pJN6. Two differences in nucleotides were found as compared with the sequences already published. One of the differences occurs at nucleotide number 754 where T replaces C. Another difference occurs at nucleotide number 1654 where A replaces G. Both substitutions do not change their amino acid codes. In order to further study the application of the cDNA as probe for RFLP at the LDL receptor locus,the probe was prepared from the recombinant plasmid and then hybridized to total genomic DNA digested with Stu I. The test confirmed that Stu I in exon 8 was a polymorphic site.
出处
《生物化学杂志》
CAS
CSCD
1996年第5期535-539,共5页
