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以茶树龙井43品种的后期花蕾为试验材料,提取总RNA,然后进行RT-PCR得到cDNA第1链。同时,根据GenBank上登录的茶树花粉特异蛋白基因CsPSP(DQ887753)的序列,设计1对特异引物进行PCR反应。将扩增后的基因片段插入pBI121表达载体中,并测序。结果表明,插入pBI121载体的片段长度为318 bp,与已知序列进行比对后一致性达到99.02%,说明载体构建成功。
Taking the late flower bud of Longjing 43 variety of tea tree as the experimental material, total RNA was extracted, and then the first strand cDNA was obtained by RT-PCR. In the meantime, a pair of specific primers was designed for PCR reaction according to the sequence of CsPSP (DQ887753) of the tea plant pollen-specific protein registered in GenBank. The amplified gene fragment was inserted into pBI121 expression vector and sequenced. The results showed that the fragment inserted into pBI121 vector was 318 bp in length and its identity with the known sequence was 99.02%, indicating that the vector was successfully constructed.