Cloning And Sequence Analysis And Expression Analysis Of Fragment Of Heat Shock Protein 70 Gene Of Locusta Migratoria Tibetensis Chen

Locusta migratoria tibetensis Chen is one of the three migratory locust subspecies in China, mainly damage the crops like corn and highland barley, distributed in Tibet, Qinghai and West Sichuan Plateau with the elevation of about 1130～4600 meters, Climate characterized by the variations of altitude, strong ultraviolet radiation and larger temperature difference between day and night. L.m.tibetensis can establish population in plateau was the result from long-term adaption of low temperature and uv radiation. The recent study of L.m.tibetensis mainly focuses on the aspects as morphology, biological characteristics and cold tolerance, but there was little-report about it’s molecular biology characteristics. In this article, we studied the inducible expression of heat shock protein 70(HSP70) by different temperature and uv radiation and cloned the core fragment of HSP70 gene.The primary contents are as follows:1. Cloned the core fragment of HSP70 gene, which is designated as Lmthsp70(Genbank accession number KU726093). This sequence contains two signature sequences of HSP70 family, who is high similar with Locusta migratoria manilensis Meyen. Our derivation of the amino acid sequence have a startling likeness to Locusta migratoria manilensis Meyen and other HSP70 amino acid sequence. It’s still has some differences, through comparison, we found that the bases of Locusta migratoria.tibetensis Hsp70 appears substitution, addition and deletion.It may because L.m.tibetensis living in constant at high altitude to get acclimatized to plateau climate.2. Cloned the fragment sepuence of β-actin gene and given names as LmtBa((Submitted)),The homology between it’s nucleotide sequence and amino acid sequence with other aspect were discovered.The sequence of mRNA of β-actin homology of 99% with Locusta migratoria manilensis Meyen(AY344445.1),the homology of 92% with Schistocerca gregaria (HQ851398.1)3. Clear the expression pattern of Tibet locust Lmthsp70 radiation at different temperatures and under UV stress.40 and 45 degrees Celsius heat shock of female adults of 1H, Lmthsp70 relative expression quantity is 20 DEG C were raised 2.3 times and 24.7-fold; 0 DEG C and-5 DEG C chilled female adult 1H, Lmthsp70 relative expression quantity is 20 DEG C, respectively, up 15.4 and 16.6 times.40 degrees C and 45 C heat stress male adult 1H, Lmthsp70 expression was 26.8 times and 25.2 times the amount of treatment were up 20 times; 0 and -5 C cold induced male adult 1H, Lmthsp70 expression levels were 15.2 times and 17.4 times respectively. In the UV-B and 30W ultraviolet radiation treatment within 60 min, Locusta migratoria manilensis Lmthsp70 gene relative expression quantity with time extended. The expression of up-regulated. The results show that Lmthsp70 is an inducible protein, is high, induced by low temperature and UV radiation, and is one of the important factor of the extreme environment of Locusta migratoria manilensis response to the plateau.